Choline salt forms of an HIV capsid inhibitor

ABSTRACT

The present disclosure relates to choline salts, and crystalline forms thereof, of a compound which is N—((S)-1-(3-(4-chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3- (trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide, which is useful in the treatment and prevention of a Retroviridae viral infection including an infection caused by the HIV virus.

TECHNICAL FIELD

The present disclosure relates to choline salt forms of an HIV capsidinhibitor, and pharmaceutical compositions thereof, for use in thetreatment or prevention of a Retroviridae viral infection including aninfection caused by the HIV virus.

BACKGROUND

Positive-single stranded RNA viruses comprising the Retroviridae familyinclude those of the subfamily Orthoretrovirinae and generaAlpharetrovirus, Betaretrovirus, Gammaretrovirus, Deltaretrovirus,Epsilonretrovirus, Lentivirus, and Spumavirus which cause many human andanimal diseases. Among the Lentivirus, HIV-1 infection in humans leadsto depletion of T helper cells and immune dysfunction, producingimmunodeficiency and vulnerability to opportunistic infections. TreatingHIV-1 infections with highly active antiretroviral therapies (HAART) hasproven to be effective at reducing viral load and significantly delayingdisease progression (Hammer, S. M., et al.; JAMA 2008, 300: 555-570).However, these treatments could lead to the emergence of HIV strainsthat are resistant to current therapies (Taiwo, B., InternationalJournal of Infectious Diseases 2009, 13:552-559; Smith, R. J., et al.,Science 2010, 327:697-701). Therefore, there is an ongoing need todiscover new antiretroviral agents and to develop methods for theirpreparation and purification as well as prepare improved pharmaceuticalformulations of the same. The choline salt forms of the HIV capsidinhibitor disclosed herein help meet these and other needs.

SUMMARY

The present application provides a choline salt of Compound 1:

The present application further provides a pharmaceutical compositioncomprising a choline salt of Compound 1, or a crystalline form thereofdisclosed herein, and at least one pharmaceutically acceptableexcipient.

The present application further provides a crystalline form of thecholine salt of Compound 1.

The present application further provides a method of treating orpreventing a human immunodeficiency virus (HIV) infection comprisingadministering a therapeutically effective amount of a choline salt, orcrystalline form thereof, to a subject in need thereof.

The present application further provides a choline salt of Compound 1,or crystalline form thereof, for use in therapy.

The present application further provides a choline salt of Compound 1,or crystalline form thereof, for use in a method of treating orpreventing a human immunodeficiency virus (HIV) infection, comprisingadministering a therapeutically effective amount of the choline salt, orcrystalline form, to a subject in need thereof.

The present application further provides a process of increasing theamount of an isomeric compound of Isomer A:

relative to an amount of an isomeric compound of Isomer B:

or the amount of an isomeric compound of Isomer B relative to the amountof an isomeric compound of Isomer A, in a starting mixture comprisingboth isomeric compounds, the process comprising:

contacting the starting mixture with N,N,N-trimethylethanolammoniumhydroxide in the presence of a solvent to form aN,N,N-trimethylethanolammonium salt mixture of both isomeric compounds,wherein the salt mixture has an increased amount of the isomeric salt ofIsomer A relative to the amount of the isomeric salt of Isomer B, or anincreased amount of the isomeric salt of Isomer B relative to the amountof the isomeric salt of Isomer A, when compared with the relativeamounts of the isomeric compounds of Isomer A and Isomer B in thestarting mixture.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows ¹H-NMR spectra characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, enriched inIsomer B (top trace) and enriched in Isomer A (bottom trace).

FIG. 2 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1 fromisopropyl alcohol (top trace: bulk crystals; bottom trace: needles).

FIG. 3 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, Material A.

FIG. 4 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm I.

FIG. 5 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm I.

FIG. 6 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm II.

FIG. 7 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm II.

FIG. 8 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm III.

FIG. 9 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm III.

FIG. 10 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm IV.

FIG. 11 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm IV.

FIG. 12 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm V.

FIG. 13 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm V.

FIG. 14 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm VI.

FIG. 15 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm VI.

FIG. 16 shows an XRPD pattern characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm VII.

FIG. 17 shows a DSC thermogram characteristic of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm VII.

FIG. 18 shows an XRPD pattern characteristic of the ethanol solvate formof the N,N,N-trimethylethanolammonium (choline) salt of Compound 1.

FIG. 19 shows an XRPD pattern characteristic of the tetrahydrofuransolvate form of the N,N,N-trimethylethanolammonium (choline) salt ofCompound 1.

FIG. 20 shows an XRPD pattern characteristic of the methyl tert-butylether solvate form of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1.

DETAILED DESCRIPTION

The present invention relates to new solid forms, which are cholinesalts, of the HIV capsid inhibitorN—((S)-1-(3-(4-chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide(i.e., Compound 1, see below). One skilled in the art understands that acompound structure may be named or identified using commonly recognizednomenclature systems and symbols. By way of example, the compound may benamed or identified with common names, systematic or non-systematicnames. The nomenclature systems and symbols that are commonly recognizedin the art of chemistry including but not limited to Chemical AbstractService (CAS) and International Union of Pure and Applied Chemistry(IUPAC). Accordingly, the compound structure for Compound 1 providedabove may also be named or identified asN—((S)-1-(3-(4-chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide.

Compound 1 can be a weak acid (pKA 6.7 at sulfonamide) or a weak base(pKa <1 at pyridine) and consists of two atropisomers, Isomer A andIsomer B, that can rotate along one of the C—C bonds, as shown below. Insolution, the two atropisomers coexist in the ratio of about 1:5 to 1:8(Isomer A:Isomer B), depending on temperatures and pH. The twoatropisomers can be separated by chromatography but they re-equilibratein solution (t_(1/2)≈-1-2 hours at 37° C. and the rotational energybarrier is about 24 kcal/mol).

As described herein, Isomer A or Isomer B can be enriched bycrystallization. For the N,N,N-trimethylethanolammonium (choline) saltsdescribed herein, either of the atropisomers can be enriched through theformation of different crystalline forms.

The solid forms of the invention include salt forms (both amorphous andcrystalline) of Compound 1. As used herein, “solid form” generallyrefers to a solid chemical substance that can be amorphous orcrystalline. In some embodiments, the solid form of the invention is acholine salt of Compound 1 which can be amorphous or crystalline.Crystalline choline salts of Compound 1 can exist in differentcrystalline forms (i.e., have different polymorphic or pseudopolymorphicforms).

As used herein, “crystalline form” is meant to refer to a certainlattice configuration of a crystalline substance (e.g., a salt).Different crystalline forms of the same substance typically havedifferent crystalline lattices (e.g., unit cells) which are attributedto different physical properties that are characteristic of each of thecrystalline forms. In some instances, different lattice configurationshave different water or solvent content.

According to the present invention, a crystalline form of a choline saltof Compound 1 can be useful in the synthesis and/or purification ofCompound 1. For example, a crystalline form of a choline salt ofCompound 1 can be an intermediate in the synthesis of Compound 1. Inaddition, different crystalline forms of choline salts of Compound 1 mayhave different properties with respect to bioavailability, stability,purity, and/or manufacturability for medical or pharmaceutical uses.Variations in the crystal structure of a pharmaceutical drug substanceor active ingredient may affect the dissolution rate (which may affectbioavailability, etc.), manufacturability (e.g., ease of handling,ability to consistently prepare doses of known strength), and stability(e.g., thermal stability, shelf life, etc.) of a pharmaceutical drugproduct or active ingredient. Such variations may affect the preparationor formulation of pharmaceutical compositions in different dosage ordelivery forms, such as solutions or solid oral dosage form includingtablets and capsules. Compared to other forms such as non-crystalline oramorphous forms, crystalline forms may provide desired or suitablehygroscopicity, particle size controls, dissolution rate, solubility,purity, physical and chemical stability, manufacturability, yield,and/or process control. Thus, the crystalline forms of the choline saltsof Compound 1 may provide advantages such as improving the manufacturingprocess of the compound, the stability or storability of a drug productform of the compound, the stability or storability of a drug substanceof the compound and/or the bioavailability and/or stability of thecompound as an active agent.

The use of certain solvents and/or processes have been found to producedifferent crystalline forms of the choline salts of Compound 1 which mayexhibit one or more of the favorable characteristics described above.The processes for the preparation of the crystalline forms describedherein and characterization of these crystalline forms are described indetail below.

In some embodiments, the choline salts described herein, or crystallineforms thereof, are purified or substantially isolated. By “substantiallyisolated” is meant that the choline salt, or crystalline form thereof isat least partially or substantially separated from the environment inwhich it was formed or detected. Partial separation can include, forexample, a composition enriched in the choline salt, or crystalline formof the invention. Substantial separation can include compositionscontaining at least about 50%, at least about 60%, at least about 70%,at least about 80%, at least about 90%, at least about 95%, at leastabout 97%, or at least about 99% by weight of the choline salt, orcrystalline form of the invention. In some embodiments, the cholinesalt, or crystalline form of the invention can be prepared with a purityof about 75% or more, 80% or more, 85% or more, 90% or more, 95% ormore, 98% or more, or 99% or more.

The different crystalline forms can be identified by solid statecharacterization methods such as by X-ray powder diffraction (XRPD).Other characterization methods such as differential scanning calorimetry(DSC) further help identify the form as well as help determine stabilityand solvent/water content.

An XRPD pattern of reflections (peaks) is typically considered afingerprint of a particular crystalline form. It is well known that therelative intensities of the XRPD peaks can widely vary depending on,inter alia, the sample preparation technique, crystal size distribution,various filters used, the sample mounting procedure, and the particularinstrument employed. In some instances, new peaks may be observed orexisting peaks may disappear, depending on the type of the instrument orthe settings. As used herein, the term “peak” refers to a reflectionhaving a relative height/intensity of at least about 5% of the maximumpeak height/intensity. Moreover, instrument variation and other factorscan affect the 2-theta values. Thus, peak assignments, such as thosereported herein, can vary by plus or minus about 0.2° (2-theta), and theterm “substantially” and “about” as used in the context of XRPD hereinis meant to encompass the above-mentioned variations.

In the same way, temperature readings in connection with DSC can varyabout ±3° C. depending on the instrument, particular settings, samplepreparation, etc. Accordingly, a crystalline form reported herein havinga DSC thermogram “substantially” as shown in any of the Figures or theterm “about” is understood to accommodate such variation.

The present invention provides crystalline forms of certain compounds orsalts thereof. In some embodiments, the crystalline form may besubstantially anhydrous. In some embodiments, the crystalline form maybe hydrated or solvated.

Compound 1 N,N,N-trimethylethanolammonium (choline) salt

In some embodiments, Compound 1 can be isolated as aN,N,N-trimethylethanolammonium (choline) salt which can be amorphous orcrystalline. In some embodiments, the N,N,N-trimethylethanolammonium(choline) salt of Compound 1 is crystalline.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1 is selected fromcrystalline Material A, crystalline Form I, crystalline Form II,crystalline Form III, crystalline Form IV, crystalline Form V,crystalline Form VI, and crystalline Form VII.

In some embodiments, crystalline Material A of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 3.

In some embodiments, crystalline Form I of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 4.

In some embodiments, crystalline Form I of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 5.5°, 7.5°, 7.9°, 14.9°, 15.7°, 16.8°,17.6°, 19.3°, and 22.4°.

In some embodiments, crystalline Form I of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 5.

In some embodiments, crystalline Form I of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 157° C.

In some embodiments, crystalline Form II of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 6.

In some embodiments, crystalline Form II of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 7.5°, 9.6°, 14.0°, 14.9°, 16.1°, 16.9°,20.8°, 21.0°, and 26.5°.

In some embodiments, crystalline Form II of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 7.

In some embodiments, crystalline Form II of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 147° C.

In some embodiments, crystalline Form III of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 8.

In some embodiments, crystalline Form III of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 7.8°, 8.1°, 8.3°, 15.0°, 15.7°, 16.7°,20.0°, 21.1°, and 21.7°.

In some embodiments, crystalline Form III of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 9.

In some embodiments, crystalline Form III of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 144° C.

In some embodiments, crystalline Form IV of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 10.

In some embodiments, crystalline Form IV of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 7.5°, 8.0°, 14.8°, 16.1°, 17.0°, 20.3°,21.1°, 24.6°, and 26.7°.

In some embodiments, crystalline Form IV of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 11.

In some embodiments, crystalline Form IV of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 136° C.

In some embodiments, crystalline Form V of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 12.

In some embodiments, crystalline Form V of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 6.9°, 7.9°, 10.7°, 16.7°, 17.6°, 21.1°,21.8°, 22.8°, and 26.9°.

In some embodiments, crystalline Form V of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 13.

In some embodiments, crystalline Form V of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 159° C.

In some embodiments, crystalline Form VI of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 14.

In some embodiments, crystalline Form VI of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 6.1°, 8.6°, 9.5°, 15.4°, 20.4°, 21.9°,22.5°, 24.2°, and 25.2°.

In some embodiments, crystalline Form VI of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 15.

In some embodiments, crystalline Form VI of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 121° C.

In some embodiments, crystalline Form VII of theN,N,N-trimethylethanolammonium salt of Compound 1 has an XRPD profilesubstantially as shown in FIG. 16.

In some embodiments, crystalline Form VII of theN,N,N-trimethylethanolammonium salt of Compound 1 has at least one, atleast two, at least three, at least four, at least five, at least six,at least seven, at least eight, or at least nine XRPD peaks, in terms of2-theta±0.2°, selected from 4.7°, 7.3°, 8.9°, 9.5°, 18.3°, 20.5°, 22.3°,24.9°, and 28.4°.

In some embodiments, crystalline Form VII of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram substantially as shown in FIG. 17.

In some embodiments, crystalline Form VII of theN,N,N-trimethylethanolammonium salt of Compound 1 is characterized by aDSC thermogram having a melting onset of about 144° C.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1 is solvated.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1 is an ethanol solvate.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1, ethanol solvate hasan XRPD profile substantially as shown in FIG. 18.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1 is a tetrahydrofuransolvate.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1, tetrahydrofuransolvate has an XRPD profile substantially as shown in FIG. 19.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1 is a methyl tert-butylether solvate.

In some embodiments, the crystalline form of theN,N,N-trimethylethanolammonium salt of Compound 1, methyl tert-butylether solvate has an XRPD profile substantially as shown in FIG. 20.

The present application further provides a process of increasing theamount of an isomeric compound of Isomer A:

relative to an amount of an isomeric compound of Isomer B:

or the amount of an isomeric compound of Isomer B relative to the amountof an isomeric compound of Isomer A, in a starting mixture comprisingboth isomeric compounds, the process comprising:

contacting the starting mixture with N,N,N-trimethylethanolammoniumhydroxide in the presence of a solvent to form aN,N,N-trimethylethanolammonium salt mixture of both isomeric compounds,wherein the salt mixture has an increased amount of the isomeric salt ofIsomer A relative to the amount of the isomeric salt of Isomer B, or anincreased amount of the isomeric salt of Isomer B relative to the amountof the isomeric salt of Isomer A, when compared with the relativeamounts of the isomeric compounds of Isomer A and Isomer B in thestarting mixture.

In some embodiments, the process comprises increasing the amount of anisomeric compound of Isomer A relative to an amount of an isomericcompound of Isomer B. In some embodiments, the solvent is selected fromthe group consisting of acetonitrile, 2-methyltetrahydrofuran,isopropylacetate, ethanol, isopropanol, tetrahydrofuran, methyltert-butyl ether, and isopropyl ether, or any mixture thereof. In someembodiments, the process of increasing the amount of an isomericcompound of Isomer A relative to an amount of an isomeric compound ofIsomer B further comprises drying the salt mixture to form a second saltmixture comprising an increased amount of an isomeric compound of IsomerA relative to an amount of an isomeric compound of Isomer B.

In some embodiments, the process comprises increasing the amount of anisomeric compound of Isomer B relative to an amount of an isomericcompound of Isomer A. In some embodiments, the solvent is selected fromthe group consisting of methanol, isopropanol, dichloromethane,isopropyl ether, heptane, and toluene, or any mixture thereof.

In some embodiments, the choline salt, or crystalline form thereofdisclosed herein is enriched in Isomer A. In some embodiments, thecholine salt or crystalline form disclosed herein is enriched in IsomerB. As used herein, the term “enriched,” refers to an increased amount ofa particular compound, salt, or isomeric compound in a mixture whencompared with the amount of the compound, salt, or isomeric compound inthe mixture prior to being enriched. For example, a mixture enriched inan isomeric compound of Isomer A has an increased amount of the isomericcompound of Isomer A relative to the isomeric compound of Isomer B whencompared with the relative amounts of the isomeric compounds of Isomer Aand Isomer B in a starting mixture.

The description below is made with the understanding that the presentdisclosure is to be considered as an exemplification of the claimedsubject matter, and is not intended to limit the appended claims to thespecific embodiments illustrated. The headings used throughout thisdisclosure are provided for convenience and are not to be construed tolimit the claims in any way. Embodiments illustrated under any headingmay be combined with embodiments illustrated under any other heading.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art.

When trade names are used herein, it is intended to independentlyinclude the tradename product and the active pharmaceuticalingredient(s) of the tradename product.

As used herein and in the appended claims, the singular forms “a” and“an”, and “the” include plural referents unless the context clearlydictates otherwise. Thus, e.g., reference to “the compound” includes aplurality of such compounds and reference to “the assay” includesreference to one or more assays, and so forth.

“Pharmaceutically acceptable” refers to compounds, salts, compositions,dosage forms and other materials which are useful in preparing apharmaceutical composition that is suitable for veterinary or humanpharmaceutical use.

“Pharmaceutically acceptable excipient” includes without limitation anyadjuvant, carrier, excipient, glidant, sweetening agent, diluent,preservative, dye/colorant, flavor enhancer, surfactant, wetting agent,dispersing agent, suspending agent, stabilizer, isotonic agent, solvent,or emulsifier which has been approved by the United States Food and DrugAdministration as being acceptable for use in humans or domesticanimals.

“Pharmaceutically acceptable salt” refers to a salt of a compound thatis pharmaceutically acceptable and that possesses (or can be convertedto a form that possesses) the desired pharmacological activity of theparent compound.

“Subject” and “subjects” refers to humans, domestic animals (e.g., dogsand cats), farm animals (e.g., cattle, horses, sheep, goats and pigs),laboratory animals (e.g., mice, rats, hamsters, guinea pigs, pigs,rabbits, dogs, and monkeys), and the like.

As used herein, “treatment” or “treating” is an approach for obtainingbeneficial or desired results. For purposes of the present disclosure,beneficial or desired results include, but are not limited to,alleviation of a symptom and/or diminishment of the extent of a symptomand/or preventing a worsening of a symptom associated with a disease orcondition. In one embodiment, “treatment” or “treating” includes one ormore of the following: a) inhibiting the disease or condition (e.g.,decreasing one or more symptoms resulting from the disease or condition,and/or diminishing the extent of the disease or condition); b) slowingor arresting the development of one or more symptoms associated with thedisease or condition (e.g., stabilizing the disease or condition,delaying the worsening or progression of the disease or condition);and/or c) relieving the disease or condition, e.g., causing theregression of clinical symptoms, ameliorating the disease state,delaying the progression of the disease, increasing the quality of life,and/or prolonging survival.

As used herein, “delaying” development of a disease or condition meansto defer, hinder, slow, retard, stabilize and/or postpone development ofthe disease or condition. This delay can be of varying lengths of time,depending on the history of the disease and/or subject being treated. Asis evident to one skilled in the art, a sufficient or significant delaycan, in effect, encompass prevention, in that the subject does notdevelop the disease or condition. For example, a method that “delays”development of AIDS is a method that reduces the probability of diseasedevelopment in a given time frame and/or reduces extent of the diseasein a given time frame, when compared to not using the method. Suchcomparisons may be based on clinical studies, using a statisticallysignificant number of subjects. For example, the development of AIDS canbe detected using known methods, such as confirming a subject's HIV+status and assessing the subject's T-cell count or other indication ofAIDS development, such as extreme fatigue, weight loss, persistentdiarrhea, high fever, swollen lymph nodes in the neck, armpits or groin,or presence of an opportunistic condition that is known to be associatedwith AIDS (e.g., a condition that is generally not present in subjectswith functioning immune systems but does occur in AIDS patients).Development may also refer to disease progression that may be initiallyundetectable and includes occurrence, recurrence and onset.

As used herein, “prevention” or “preventing” refers to a regimen thatprotects against the onset of the disease or disorder such that theclinical symptoms of the disease do not develop. Thus, “prevention”relates to administration of a therapy (e.g., administration of atherapeutic substance) to a subject before signs of the disease aredetectable in the subject (e.g., administration of a therapeuticsubstance to a subject in the absence of detectable infectious agent(e.g., virus) in the subject). The subject may be an individual at riskof developing the disease or disorder, such as an individual who has oneor more risk factors known to be associated with development or onset ofthe disease or disorder. Thus, the term “preventing HIV infection”refers to administering to a subject who does not have a detectable HIVinfection an anti-HIV therapeutic substance. It is understood that thesubject for anti-HIV preventative therapy may be an individual at riskof contracting the HIV virus. Further, it is understood that preventionmay not result in complete protection against onset of the disease ordisorder. In some instances, prevention includes reducing the risk ofdeveloping the disease or disorder. The reduction of the risk may notresult in complete elimination of the risk of developing the disease ordisorder.

As used herein, an “at risk” individual is an individual who is at riskof developing a condition to be treated. An individual “at risk” may ormay not have detectable disease or condition, and may or may not havedisplayed detectable disease prior to the treatment of methods describedherein. “At risk” denotes that an individual has one or more so-calledrisk factors, which are measurable parameters that correlate withdevelopment of a disease or condition and are known in the art. Anindividual having one or more of these risk factors has a higherprobability of developing the disease or condition than an individualwithout these risk factor(s). For example, individuals at risk for AIDSare those having HIV.

As used herein, the term “therapeutically effective amount” or“effective amount” refers to an amount that is effective to elicit thedesired biological or medical response, including the amount of acompound that, when administered to a subject for treating a disease, issufficient to effect such treatment for the disease or to an amount thatis effective to protect against the contracting or onset of a disease.The effective amount will vary depending on the compound, the disease,and its severity and the age, weight, etc., of the subject to betreated. The effective amount can include a range of amounts. As isunderstood in the art, an effective amount may be in one or more doses,i.e., a single dose or multiple doses may be required to achieve thedesired treatment outcome. An effective amount may be considered in thecontext of administering one or more therapeutic agents, and a singleagent may be considered to be given in an effective amount if, inconjunction with one or more other agents, a desirable or beneficialresult may be or is achieved. Suitable doses of any co-administeredcompounds may optionally be lowered due to the combined action (e.g.,additive or synergistic effects) of the compounds.

“Enantiomers” are a pair of stereoisomers that are non-superimposablemirror images of each other. A 1:1 mixture of a pair of enantiomers is a“racemic” mixture. A mixture of enantiomers at a ratio other than 1:1 isa “scalemic” mixture.

“Diastereoisomers” are stereoisomers that have at least two asymmetricatoms, but which are not mirror-images of each other.

The absolute stereochemistry is specified according to theCahn-Ingold-Prelog R—S system. When a compound is a pure enantiomer thestereochemistry at each chiral carbon may be specified by either R or S.Resolved compounds whose absolute configuration is unknown can bedesignated (+) or (−) depending on the direction (dextro- orlevorotatory) which they rotate plane polarized light at the wavelengthof the sodium D line. Certain of the compounds and salts describedherein contain one or more asymmetric centers and/or hindered rotationabout a bond axis and may thus give rise to enantiomers, diastereomers,and other stereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)— or (S)—. The present disclosure is meant toinclude all such possible isomers, including racemic mixtures, scalemicmixtures, diastereomeric mixtures, optically pure forms and intermediatemixtures. Optically active (R)- and (S)-isomers may be prepared usingchiral synthons or chiral reagents, or resolved using conventionaltechniques.

Except as expressly defined otherwise, the present disclosure includesall tautomers of compounds detailed herein, even if only one tautomer isexpressly represented (e.g., both tautomeric forms are intended anddescribed by the presentation of one tautomeric form where a pair of twotautomers may exist). For example, if reference is made to a compoundcontaining an amide (e.g., by structure or chemical name), it isunderstood that the corresponding imidic acid tautomer is included bythis disclosure and described the same as if the amide were expresslyrecited either alone or together with the imidic acid. Where more thantwo tautomers may exist, the present disclosure includes all suchtautomers even if only a single tautomeric form is depicted by chemicalname and/or structure.

It is understood by one skilled in the art that this disclosure alsoincludes any salt disclosed herein may be enriched at any or all atomsabove naturally occurring isotopic ratios with one or more isotopes suchas, but not limited to, deuterium (²H or D).

Disclosed are also choline salts of Compound 1 in which from 1 to nhydrogen atoms attached to a carbon atom may be replaced by a deuteriumatom or D, in which n is the number of hydrogen atoms in the molecule.As known in the art, the deuterium atom is a non-radioactive isotope ofthe hydrogen atom. Such salts may increase resistance to metabolism, andthus may be useful for increasing the half-life of the compounds whenadministered to a mammal. See, e.g., Foster, “Deuterium Isotope Effectsin Studies of Drug Metabolism”, Trends Pharmacol. Sci., 5(12):524-527(1984). Such salts are synthesized by means well known in the art, forexample by employing starting materials in which one or more hydrogenatoms have been replaced by deuterium.

Examples of isotopes that can be incorporated into the disclosed saltsalso include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, chlorine, and iodine, such as ²H, ³H, ¹¹C, ¹³C,¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I,respectively. Substitution with positron emitting isotopes, such as ¹¹,¹⁸F, ¹⁵O and ¹³N, can be useful in Positron Emission Topography (PET)studies for examining substrate receptor occupancy. Isotopically-labeledsalts can generally be prepared by conventional techniques known tothose skilled in the art or by processes analogous to a lower predictedhepatic clearance relative to compounds A and B (about 9 and about 4time reagent in place of the non-labeled reagent previously employed.

Compounds described herein may have chiral centers and/or geometricisomeric centers (E- and Z-isomers), and it is to be understood that allsuch optical, enantiomeric, diastereoisomeric and geometric isomers areencompassed. Where compounds are represented in their chiral form, it isunderstood that the embodiment encompasses, but is not limited to, thespecific diastereomerically or enantiomerically enriched form. Wherechirality is not specified but is present, it is understood that theembodiment is directed to either the specific diastereomerically orenantiomerically enriched form; or a racemic or scalemic mixture of suchcompound(s).

In a preferred embodiment, the current disclosure relates to the use ofthe choline salts and crystalline forms of the invention in treating aRetroviridae viral infection including an infection caused by the HIVvirus comprising administering a therapeutically effective amount of thecholine salt or crystalline form to a subject in need thereof.

In a preferred embodiment, the current disclosure relates to the use ofthe choline salts and crystalline forms of the invention in treating aRetroviridae viral infection including an infection caused by the HIVvirus comprising administering a therapeutically effective amount of thecholine salt or crystalline form to a subject in need thereof.

It is a desirable goal to discover a compound, a choline salt, orcrystalline form thereof having a low EC₅₀. The EC₅₀ value refers to theconcentration of a compound in an assay that achieves 50% of the maximumefficacy. A compound, salt, or crystalline form with a lower EC₅₀achieves similar efficacy with lower compound, salt, or crystalline formconcentration relative to a compound, salt, or crystalline form with ahigher EC₅₀. Thus, a lower EC₅₀ is generally preferred for drugdevelopment.

It is a desirable goal to discover a compound, pharmaceuticallyacceptable salt, or crystalline form thereof that has good physicaland/or chemical stability. An increase in overall stability of acompound, salt, or crystalline form can provide an increase incirculation time in the body. With less degradation, a stable compound,salt, or crystalline form can be administered in lower doses and stillmaintain efficacy. Also, with less degradation, there is less concernabout by-products from degradation of a compound or salt.

It is a desirable goal to discover a compound, pharmaceuticallyacceptable salt, or crystalline form thereof that has improvedpharmacokinetic and/or pharmacodynamic profiles and long half-life. Itis advantageous for a drug to have a moderate or low clearance and along half-life, as this can lead to a good bioavailability and highexposure in systemic exposure. Reducing the clearance and increasinghalf-life time of a compound, salt, or crystalline form could reduce thedaily dose required for efficacy and therefore give a better efficacyand safety profile. Thus, improved pharmacokinetic and/orpharmacodynamic profiles and long half-life can provide for betterpatient compliance.

It is a desirable goal to discover a compound, a pharmaceuticallyacceptable salt, or crystalline form thereof that has goodpharmacokinetic profile from a slow release injectable formulation. Itis advantageous for a drug to have a low EC₅₀ and long actingpharmacokinetics, as this can lead to low frequency of administration.Reducing the frequency of administration can provide for better patientcompliance. Reducing the frequency of administration can be desirablefor patients with difficult or limited access to health care.

Methods of Use

In some embodiments, the choline salts or crystalline forms disclosedherein are used for preventing an HIV infection in a subject. In someembodiments, the choline salts or crystalline forms disclosed herein areused for preventing an HIV infection in a subject at risk for infection.In some embodiments, the choline salts or crystalline forms disclosedherein are used for pre-exposure prophylaxis (PrEP) to reduce the riskof sexually acquired HIV-1. It is believed that the pharmaceuticallyaccpetable salts or crystalline forms disclosed herein are activeagainst major HIV-1 mutants selected by clinical Protease Inhibitors(PIs), nucleoside reverse transcriptase inhibitors (NRTIs),Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs), and Integraseinhibitors (INSTIs).

In certain embodiments, a method for treating or preventing an HIVinfection in a subject (e.g., a human), comprising administering acholine salt of Compound 1, or crystalline form thereof, to the subjectis disclosed.

In some embodiments, a method for treating or preventing an HIVinfection in a subject (e.g., a human), comprising administering acholine salt of Compound 1, or crystalline form thereof, to the subjectis disclosed.

In certain embodiments, a method for inhibiting the replication of theHIV virus, treating AIDS or delaying the onset of AIDS in a subject(e.g., a human), comprising administering a choline salt of Compound 1,or crystalline form thereof, to the subject is disclosed.

In some embodiments, a method for inhibiting the replication of the HIVvirus, treating AIDS or delaying the onset of AIDS in a subject (e.g., ahuman), comprising administering a choline salt of Compound 1, orcrystalline form thereof, to the subject is disclosed.

In certain embodiments, a method for preventing an HIV infection in asubject (e.g., a human), comprising administering a choline salt ofCompound 1, or crystalline form thereof, to the subject is disclosed. Incertain embodiments, the subject is at risk of contracting the HIVvirus, such as a subject who has one or more risk factors known to beassociated with contracting the HIV virus.

In some embodiments, a method for preventing an HIV infection in asubject (e.g., a human), comprising administering a therapeuticallyeffective amount of a choline salt of Compound 1, or crystalline formthereof, to the subject is disclosed. In certain embodiments, thesubject is at risk of contracting the HIV virus, such as a subject whohas one or more risk factors known to be associated with contracting theHIV virus.

In certain embodiments, a method for treating an HIV infection in asubject (e.g., a human), comprising administering a choline salt ofCompound 1, or crystalline form thereof, to the subject is disclosed.

In some embodiments, a method for treating an HIV infection in a subject(e.g., a human), comprising administering a choline salt of Compound 1,or crystalline form thereof, to the subject is disclosed.

In certain embodiments, a method for treating an HIV infection in asubject (e.g., a human), comprising administering to the subject in needthereof a therapeutically effective amount of a choline salt of Compound1, or crystalline form thereof, in combination with a therapeuticallyeffective amount of one or more (e.g., one, two, three, or four; or oneor two; or one to three; or one to four) additional therapeutic agentsselected from the group consisting of HIV protease inhibiting compounds,HIV non-nucleoside inhibitors of reverse transcriptase, HIVnon-nucleotide inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4inhibitors, gp120 inhibitors, CCR5 inhibitors, capsid polymerizationinhibitors, pharmacokinetic enhancers, and other drugs for treating HIV,and combinations thereof is disclosed. In certain embodiments, a methodfor treating an HIV infection in a subject (e.g., a human), comprisingadministering to the subject in need thereof a therapeutically effectiveamount of a choline salt of Compound 1, or crystalline form thereof, incombination with a therapeutically effective amount of one or more(e.g., one, two, three, or four; or one or two; or one to three; or oneto four) additional therapeutic agents selected from the groupconsisting of combination drugs for HIV, other drugs for treating HIV,HIV protease inhibitors, HIV non-nucleoside or non-nucleotide inhibitorsof reverse transcriptase, HIV nucleoside or nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site(or allosteric) integrase inhibitors, HIV entry inhibitors, HIVmaturation inhibitors, latency reversing agents, compounds that targetthe HIV capsid, immune-based therapies, phosphatidylinositol 3-kinase(PI3K) inhibitors, HIV antibodies, bispecific antibodies and“antibody-like” therapeutic proteins, HIV p17 matrix protein inhibitors,IL-13 antagonists, peptidyl-prolyl cis-trans isomerase A modulators,protein disulfide isomerase inhibitors, complement C5a receptorantagonists, DNA methyltransferase inhibitor, HIV vif gene modulators,Vif dimerization antagonists, HIV-1 viral infectivity factor inhibitors,TAT protein inhibitors, HIV-1 Nef modulators, Hck tyrosine kinasemodulators, mixed lineage kinase-3 (MLK-3) inhibitors, HIV-1 splicinginhibitors, Rev protein inhibitors, integrin antagonists, nucleoproteininhibitors, splicing factor modulators, COMM domain containing protein 1modulators, HIV ribonuclease H inhibitors, retrocyclin modulators, CDK-9inhibitors, dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAGprotein inhibitors, HIV POL protein inhibitors, Complement Factor Hmodulators, ubiquitin ligase inhibitors, deoxycytidine kinaseinhibitors, cyclin dependent kinase inhibitors, proprotein convertasePC9 stimulators, ATP dependent RNA helicase DDX3X inhibitors, reversetranscriptase priming complex inhibitors, G6PD and NADH-oxidaseinhibitors, pharmacokinetic enhancers, HIV gene therapy, and HIVvaccines, or any combinations thereof is disclosed.

In some embodiments, a method for treating an HIV infection in a subject(e.g., a human), comprising administering to the subject in need thereofa therapeutically effective amount of a choline salt of Compound 1, orcrystalline form thereof, in combination with a therapeuticallyeffective amount of one or more (e.g., one, two, three, or four; or oneor two; or one to three; or one to four) additional therapeutic agentsselected from the group consisting of HIV protease inhibiting compounds,HIV non-nucleoside inhibitors of reverse transcriptase, HIVnon-nucleotide inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4inhibitors, gp120 inhibitors, CCR5 inhibitors, capsid polymerizationinhibitors, pharmacokinetic enhancers, and other drugs for treating HIV,and combinations thereof is disclosed. In certain embodiments, a methodfor treating an HIV infection in a subject (e.g., a human), comprisingadministering to the subject in need thereof a therapeutically effectiveamount of a choline salt of Compound 1, or crystalline form thereof, incombination with a therapeutically effective amount of one or more(e.g., one, two, three, or four; or one or two; or one to three; or oneto four) additional therapeutic agents selected from the groupconsisting of combination drugs for HIV, other drugs for treating HIV,HIV protease inhibitors, HIV non-nucleoside or non-nucleotide inhibitorsof reverse transcriptase, HIV nucleoside or nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site(or allosteric) integrase inhibitors, HIV entry inhibitors, HIVmaturation inhibitors, latency reversing agents, compounds that targetthe HIV capsid, immune-based therapies, phosphatidylinositol 3-kinase(PI3K) inhibitors, HIV antibodies, bispecific antibodies and“antibody-like” therapeutic proteins, HIV p17 matrix protein inhibitors,IL-13 antagonists, peptidyl-prolyl cis-trans isomerase A modulators,protein disulfide isomerase inhibitors, complement C5a receptorantagonists, DNA methyltransferase inhibitor, HIV vif gene modulators,Vif dimerization antagonists, HIV-1 viral infectivity factor inhibitors,TAT protein inhibitors, HIV-1 Nef modulators, Hck tyrosine kinasemodulators, mixed lineage kinase-3 (MLK-3) inhibitors, HIV-1 splicinginhibitors, Rev protein inhibitors, integrin antagonists, nucleoproteininhibitors, splicing factor modulators, COMM domain containing protein 1modulators, HIV ribonuclease H inhibitors, retrocyclin modulators, CDK-9inhibitors, dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAGprotein inhibitors, HIV POL protein inhibitors, Complement Factor Hmodulators, ubiquitin ligase inhibitors, deoxycytidine kinaseinhibitors, cyclin dependent kinase inhibitors, proprotein convertasePC9 stimulators, ATP dependent RNA helicase DDX3X inhibitors, reversetranscriptase priming complex inhibitors, G6PD and NADH-oxidaseinhibitors, pharmacokinetic enhancers, HIV gene therapy, and HIVvaccines, or any combinations thereof is disclosed.

In certain embodiments, a method for treating an HIV infection in asubject (e.g., a human), comprising administering to the subject in needthereof a therapeutically effective amount of a pharmaceuticallyacceptable salt of Compound 1, or a cocrystal or crystalline formthereof, in combination with a therapeutically effective amount of oneor more (e.g., one, two, three, or four; or one or two; or one to three;or one to four) additional therapeutic agents selected from the groupconsisting of combination drugs for HIV, other drugs for treating HIV,HIV protease inhibitors, HIV non-nucleoside or non-nucleotide inhibitorsof reverse transcriptase, HIV nucleoside or nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site(or allosteric) integrase inhibitors, and HIV nucleoside reversetranscriptase translocation inhibitors.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, for use in medical therapy of an HIV infection (e.g. HIV-1or the replication of the HIV virus (e.g. HIV-1) or AIDS or delaying theonset of AIDS in a subject (e.g., a human)) is disclosed.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, for use in medical therapy of an HIV infection (e.g. HIV-1 orthe replication of the HIV virus (e.g. HIV-1) or AIDS or delaying theonset of AIDS in a subject (e.g., a human)) is disclosed.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, for use in the manufacture of a medicament for treating anHIV infection or the replication of the HIV virus or AIDS or delayingthe onset of AIDS in a subject (e.g., a human) is disclosed. Oneembodiment relates to a choline salt of Compound 1, or crystalline formthereof, for use in the prophylactic or therapeutic treatment of an HIVinfection or AIDS or for use in the therapeutic treatment or delayingthe onset of AIDS.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof for use in the manufacture of a medicament for treating an HIVinfection or the replication of the HIV virus or AIDS or delaying theonset of AIDS in a subject (e.g., a human) is disclosed. One embodimentrelates to a choline salt of Compound 1, or crystalline form thereof,for use in the prophylactic or therapeutic treatment of an HIV infectionor AIDS or for use in the therapeutic treatment or delaying the onset ofAIDS.

In certain embodiments, the use of a choline salt of Compound 1, orcrystalline form thereof, for the manufacture of a medicament for an HIVinfection in a subject (e.g., a human) is disclosed. In certainembodiments, a choline salt of Compound 1, or crystalline form thereof,for use in the prophylactic or therapeutic treatment of an HIV infectionis disclosed.

In some embodiments, the use of a choline salt of Compound 1, orcrystalline form thereof, for the manufacture of a medicament for an HIVinfection in a subject (e.g., a human) is disclosed. In certainembodiments, a choline salt of Compound 1, or crystalline form thereof,for use in the prophylactic or therapeutic treatment of an HIV infectionis disclosed.

In certain embodiments, in the methods of use, the administration is toa subject (e.g., a human) in need of the treatment. In certainembodiments, in the methods of use, the administration is to a subject(e.g., a human) who is at risk of developing AIDS.

Disclosed herein is a choline salt of Compound 1, or crystalline formthereof, for use in therapy. In one embodiment, the choline salt ofCompound 1, or crystalline form thereof, is for use in a method oftreating an HIV infection or the replication of the HIV virus or AIDS ordelaying the onset of AIDS in a subject (e.g., a human).

In some embodiments, disclosed herein is a choline salt of Compound 1,or crystalline form thereof, for use in therapy. In some embodiments,the choline salt of Compound 1, or crystalline form thereof, is for usein a method of treating an HIV infection or the replication of the HIVvirus or AIDS or delaying the onset of AIDS in a subject (e.g., ahuman).

Also disclosed herein is a choline salt of Compound 1, or crystallineform thereof, for use in a method of treating or preventing HIVinfection in a subject in need thereof. In certain embodiments, acholine salt of Compound 1, or crystalline form thereof, for use in amethod of treating HIV infection in a subject in need thereof isprovided. In certain embodiments, the subject in need thereof is a humanwho has been infected with HIV. In certain embodiments, the subject inneed thereof is a human who has been infected with HIV but who has notdeveloped AIDS. In certain embodiments, the subject in need thereof is asubject at risk for developing AIDS. In certain embodiments, the subjectin need thereof is a human who has been infected with HIV and who hasdeveloped AIDS.

In some embodiments, disclosed herein is a choline salt of Compound 1,or crystalline form thereof, for use in a method of treating orpreventing HIV infection in a subject in need thereof. In certainembodiments, a choline salt of Compound 1, or crystalline form thereof,for use in a method of treating HIV infection in a subject in needthereof is provided. In certain embodiments, the subject in need thereofis a human who has been infected with HIV. In certain embodiments, thesubject in need thereof is a human who has been infected with HIV butwho has not developed AIDS. In certain embodiments, the subject in needthereof is a subject at risk for developing AIDS. In certainembodiments, the subject in need thereof is a human who has beeninfected with HIV and who has developed AIDS.

In one embodiment, a choline salt of Compound 1, or crystalline formthereof, in combination with one or more (e.g. one, two, three, or four;or one or two; or one to three; or one to four) additional therapeuticagents as described herein for use in a method of treating or preventingHIV infection in a subject in need thereof is provided. In oneembodiment, said additional therapeutic agents are selected from thegroup consisting of combination drugs for HIV, other drugs for treatingHIV, HIV protease inhibitors, HIV non-nucleoside or non-nucleotideinhibitors of reverse transcriptase, HIV nucleoside or nucleotideinhibitors of reverse transcriptase, HIV integrase inhibitors, HIVnon-catalytic site (or allosteric) integrase inhibitors, HIV entryinhibitors, HIV maturation inhibitors, latency reversing agents,compounds that target the HIV capsid, immune-based therapies,phosphatidylinositol 3-kinase (PI3K) inhibitors, HIV antibodies,bispecific antibodies and “antibody-like” therapeutic proteins, HIV p17matrix protein inhibitors, IL-13 antagonists, peptidyl-prolyl cis-transisomerase A modulators, protein disulfide isomerase inhibitors,complement C5a receptor antagonists, DNA methyltransferase inhibitor,HIV vif gene modulators, Vif dimerization antagonists, HIV-1 viralinfectivity factor inhibitors, TAT protein inhibitors, HIV-1 Nefmodulators, Hck tyrosine kinase modulators, mixed lineage kinase-3(MLK-3) inhibitors, HIV-1 splicing inhibitors, Rev protein inhibitors,integrin antagonists, nucleoprotein inhibitors, splicing factormodulators, COMM domain containing protein 1 modulators, HIVribonuclease H inhibitors, retrocyclin modulators, CDK-9 inhibitors,dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAG proteininhibitors, HIV POL protein inhibitors, Complement Factor H modulators,ubiquitin ligase inhibitors, deoxycytidine kinase inhibitors, cyclindependent kinase inhibitors, proprotein convertase PC9 stimulators, ATPdependent RNA helicase DDX3X inhibitors, reverse transcriptase primingcomplex inhibitors, G6PD and NADH-oxidase inhibitors, pharmacokineticenhancers, HIV gene therapy, and HIV vaccines, or any combinationsthereof. In one embodiment, said additional therapeutic agents areselected from the group consisting of HIV protease inhibiting compounds,HIV non-nucleoside inhibitors of reverse transcriptase, HIVnon-nucleotide inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4inhibitors, gp120 inhibitors, CCR5 inhibitors, capsid polymerizationinhibitors, pharmacokinetic enhancers, and other drugs for treating HIV,and combinations thereof.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, in combination with one or more (e.g. one, two, three, or four;or one or two; or one to three; or one to four) additional therapeuticagents as described herein for use in a method of treating or preventingHIV infection in a subject in need thereof is provided. In oneembodiment, said additional therapeutic agents are selected from thegroup consisting of combination drugs for HIV, other drugs for treatingHIV, HIV protease inhibitors, HIV non-nucleoside or non-nucleotideinhibitors of reverse transcriptase, HIV nucleoside or nucleotideinhibitors of reverse transcriptase, HIV integrase inhibitors, HIVnon-catalytic site (or allosteric) integrase inhibitors, HIV entryinhibitors, HIV maturation inhibitors, latency reversing agents,compounds that target the HIV capsid, immune-based therapies,phosphatidylinositol 3-kinase (PI3K) inhibitors, HIV antibodies,bispecific antibodies and “antibody-like” therapeutic proteins, HIV p17matrix protein inhibitors, IL-13 antagonists, peptidyl-prolyl cis-transisomerase A modulators, protein disulfide isomerase inhibitors,complement C5a receptor antagonists, DNA methyltransferase inhibitor,HIV vif gene modulators, Vif dimerization antagonists, HIV-1 viralinfectivity factor inhibitors, TAT protein inhibitors, HIV-1 Nefmodulators, Hck tyrosine kinase modulators, mixed lineage kinase-3(MLK-3) inhibitors, HIV-1 splicing inhibitors, Rev protein inhibitors,integrin antagonists, nucleoprotein inhibitors, splicing factormodulators, COMM domain containing protein 1 modulators, HIVribonuclease H inhibitors, retrocyclin modulators, CDK-9 inhibitors,dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAG proteininhibitors, HIV POL protein inhibitors, Complement Factor H modulators,ubiquitin ligase inhibitors, deoxycytidine kinase inhibitors, cyclindependent kinase inhibitors, proprotein convertase PC9 stimulators, ATPdependent RNA helicase DDX3X inhibitors, reverse transcriptase primingcomplex inhibitors, G6PD and NADH-oxidase inhibitors, pharmacokineticenhancers, HIV gene therapy, and HIV vaccines, or any combinationsthereof. In one embodiment, said additional therapeutic agents areselected from the group consisting of HIV protease inhibiting compounds,HIV non-nucleoside inhibitors of reverse transcriptase, HIVnon-nucleotide inhibitors of reverse transcriptase, HIV nucleosideinhibitors of reverse transcriptase, HIV nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4inhibitors, gp120 inhibitors, CCR5 inhibitors, capsid polymerizationinhibitors, pharmacokinetic enhancers, and other drugs for treating HIV,and combinations thereof.

In one embodiment, a choline salt of Compound 1, or crystalline formthereof, in combination with a first additional therapeutic agentselected from the group consisting of tenofovir alafenamide fumarate,tenofovir alafenamide, and tenofovir alafenamide hemifumarate, and asecond additional therapeutic agent, wherein the second additionaltherapeutic agent is emtricitabine, is provided for use in a method oftreating or preventing HIV infection in a subject in need thereof. In aparticular embodiment, a choline salt of Compound 1, or crystalline formthereof, in combination with a first additional therapeutic agentselected from the group consisting of tenofovir disoproxil fumarate,tenofovir disoproxil, and tenofovir disoproxil hemifumarate, and asecond additional therapeutic agent, wherein the second additionaltherapeutic agent is emtricitabine, is provided for use in a method oftreating or preventing HIV infection in a subject in need thereof.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, in combination with a first additional therapeutic agentselected from the group consisting of tenofovir alafenamide fumarate,tenofovir alafenamide, and tenofovir alafenamide hemifumarate, and asecond additional therapeutic agent, wherein the second additionaltherapeutic agent is emtricitabine, is provided for use in a method oftreating or preventing HIV infection in a subject in need thereof. In aparticular embodiment, a choline salt of Compound 1, or crystalline formthereof, in combination with a first additional therapeutic agentselected from the group consisting of tenofovir disoproxil fumarate,tenofovir disoproxil, and tenofovir disoproxil hemifumarate, and asecond additional therapeutic agent, wherein the second additionaltherapeutic agent is emtricitabine, is provided for use in a method oftreating or preventing HIV infection in a subject in need thereof.

In some embodiments, a pharmaceutically acceptable salt of Compound 1,or a cocrystal or crystalline form thereof, is provided in combinationwith at least one additional therapeutic agent selected from the groupconsisting of:

-   -   (1) nucleoside reverse transcriptase translocation inhibitors        (“NRTTIs”), such as 4′-Ethynyl-2-fluoro-2′-deoxyadenosine        triphosphate (also known as MK-8591 and EFdA);    -   (2) nucleoside or nucleotide reverse-transcriptase inhibitors        (“NRTIs”), such as tenofovir alafenamide fumarate, tenofovir        alafenamide, tenofovir alafenamide hemifumarate, GS-9131, and        GS-9148;    -   (3) non-nucleoside or non-nucleotide reverse transcriptase        inhibitors (“NNRTIs”), such as efavirenz, etravirine,        rilpivirine, nevirapine, and delavirdine;    -   (4) protease Inhibitors (“PIs”), such as amprenavir, atazanavir,        brecanavir, darunavir, fosamprenavir, fosamprenavir calcium,        indinavir, indinavir sulfate, lopinavir, nelfinavir, nelfinavir        mesylate, ritonavir, saquinavir, saquinavir mesylate,        tipranavir, DG-17, TMB-657 (PPL-100), T-169, BL-008, and        TMC-31091; and    -   (5) integrase strand transfer inhibitors (“INSTIs”), such as        Bictegravir, cabotegravir, raltegravir, and dolutegravir.

In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, is provided for use to prevent HIV infection from takinghold if the individual is exposed to the virus and/or to keep the virusfrom establishing a permanent infection and/or to prevent the appearanceof symptoms of the disease and/or to prevent the virus from reachingdetectable levels in the blood, for example for pre-exposure prophylaxis(PrEP) or post-exposure prophylaxis (PEP). Accordingly, in certainembodiments, methods for reducing the risk of acquiring HIV (e.g., HIV-1and/or HIV-2) are provided. For example, methods for reducing the riskof acquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administration of acholine salt of Compound 1, or crystalline form thereof. In certainembodiments, methods for reducing the risk of acquiring HIV (e.g., HIV-1and/or HIV-2) comprise administration of a choline salt of Compound 1,or crystalline form thereof, in combination with one or more additionaltherapeutic agents. In certain embodiments, methods for reducing therisk of acquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administrationof a pharmaceutical composition comprising a therapeutically effectiveamount of a choline salt of Compound 1, or crystalline form thereof, anda choline excipient.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, is provided for use to prevent HIV infection from taking holdif the individual is exposed to the virus and/or to keep the virus fromestablishing a permanent infection and/or to prevent the appearance ofsymptoms of the disease and/or to prevent the virus from reachingdetectable levels in the blood, for example for pre-exposure prophylaxis(PrEP) or post-exposure prophylaxis (PEP). Accordingly, in certainembodiments, methods for reducing the risk of acquiring HIV (e.g., HIV-1and/or HIV-2) are provided. For example, methods for reducing the riskof acquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administration of acholine salt of Compound 1, or crystalline form thereof. In certainembodiments, methods for reducing the risk of acquiring HIV (e.g., HIV-1and/or HIV-2) comprise administration of a choline salt of Compound 1,or crystalline form thereof, in combination with one or more additionaltherapeutic agents. In certain embodiments, methods for reducing therisk of acquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administrationof a pharmaceutical composition comprising a therapeutically effectiveamount of a choline salt of Compound 1, or crystalline form thereof, anda choline excipient.

In certain embodiments, methods for reducing the risk of acquiring HIV(e.g., HIV-1 and/or HIV-2) comprise administration of a choline salt ofCompound 1, or crystalline form thereof, in combination with safer sexpractices. In certain embodiments, methods for reducing the risk ofacquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administration to anindividual at risk of acquiring HIV. Examples of individuals at highrisk for acquiring HIV include, without limitation, an individual who isat risk of sexual transmission of HIV.

In some embodiments, methods for reducing the risk of acquiring HIV(e.g., HIV-1 and/or HIV-2) comprise administration of a choline salt ofCompound 1, or crystalline form thereof, in combination with safer sexpractices. In certain embodiments, methods for reducing the risk ofacquiring HIV (e.g., HIV-1 and/or HIV-2) comprise administration to anindividual at risk of acquiring HIV. Examples of individuals at highrisk for acquiring HIV include, without limitation, an individual who isat risk of sexual transmission of HIV.

In certain embodiments, the reduction in risk of acquiring HIV is atleast about 40%, 50%, 60%, 70%, 80%, 90%, or 95%. In certainembodiments, the reduction in risk of acquiring HIV is at least about75%. In certain embodiments, the reduction in risk of acquiring HIV isabout 80%, 85%, or 90%.

In another embodiment, the use of a choline salt of Compound 1, orcrystalline form thereof, for the manufacture of a medicament for thetreatment of an HIV infection in a human being having or at risk ofhaving the infection is disclosed.

In some embodiments, the use of a choline salt of Compound 1, orcrystalline form thereof, for the manufacture of a medicament for thetreatment of an HIV infection in a human being having or at risk ofhaving the infection is disclosed.

Also disclosed herein is a choline salt of Compound 1, or crystallineform thereof, for use in the therapeutic treatment or delaying the onsetof AIDS.

In some embodiments, disclosed herein is a choline salt of Compound 1,or crystalline form thereof, for use in the therapeutic treatment ordelaying the onset of AIDS.

Also disclosed herein is a choline salt of Compound 1, or crystallineform thereof, for use in the prophylactic or therapeutic treatment of anHIV infection.

In some embodiments, disclosed herein is a choline salt of Compound 1,or crystalline form thereof, for use in the prophylactic or therapeutictreatment of an HIV infection.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, can be used as a research tool.

Routes of Administration

The choline salt of Compound 1, or crystalline form thereof, (alsoreferred to herein as the active ingredient) can be administered by anyroute appropriate to the condition to be treated. Suitable routesinclude oral, rectal, nasal, topical (including buccal and sublingual),transdermal, vaginal and parenteral (including subcutaneous,intramuscular, intravenous, intradermal, intrathecal and epidural), andthe like. It will be appreciated that the preferred route may vary with,for example, the condition of the recipient. In certain embodiments, thecompounds disclosed can be dosed parenterally. In certain embodiments,the compounds disclosed can be dosed intravenous, subcutaneous, orintramuscular. In certain embodiments, the compounds, salts, andcrystalline forms disclosed are orally bioavailable and can be dosedorally.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, may be administered with a syringe suitable for administrationof the compound. In some embodiments, the syringe is disposable. In someembodiments, the syringe is reusable. In some embodiments, the syringeis pre-filled with the choline salt of Compound 1, or crystalline formthereof.

In some embodiments, the pharmaceutically acceptable salt of Compound 1,or a cocrystal or crystalline form thereof, may be administered viainjection, using an injection device. In some embodiments, the injectiondevice is or includes a syringe, which can be employed manually, or aspart of a syringe-containing injection device, such as, but not limitedto, one with a needle safety shield. A wide variety of injection devicescan be used, such as, for example and not limited to, a handheld orwearable autoinjector, a handheld or wearable manual injector, anon-body injector, a syrette, a jet injector, or a pen injector, each ofwhich can be reusable or disposable.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, may be administered with an auto-injector comprising a syringe.In some embodiments, the syringe is disposable. In some embodiments, thesyringe is reusable. In some embodiments, the syringe is pre-filled withthe choline salt of Compound 1, or crystalline form thereof.

Dosing Regimen

The choline salt of Compound 1, or crystalline form thereof, may beadministered to a subject in accordance with an effective dosing regimenfor a desired period of time or duration, such as at least about oneday, at least about one week, at least about one month, at least about 2months, at least about 3 months, at least about 4 months, at least about6 months, or at least about 12 months or longer. In one variation, thesalt or crystalline form is administered on a daily or intermittentschedule. In one variation, the salt or crystalline form is administeredon a monthly schedule. In one variation, the salt or crystalline form isadministered every two months. In one variation, the salt or crystallineform is administered every three months. In one variation, the salt orcrystalline form is administered every four months. In one variation,the salt or crystalline form is administered every five months. In onevariation, the salt or crystalline form is administered every 6 months.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, may be administered to a subject at least about one month, atleast about 4 months, or at least about 6 months. In some embodiments,the choline salt of Compound 1, or crystalline form thereof, may besubcutaneously administered to a subject at least about one month. Insome embodiments, the choline salt of Compound 1, or crystalline formthereof, may be subcutaneously or intramuscularly administered to asubject at least about 4 months, or at least about 6 months.

The dosage or dosing frequency of the choline salt of Compound 1, orcrystalline form thereof, may be adjusted over the course of thetreatment, based on the judgment of the administering physician.

In some embodiments, the dosage or dosing frequency of the choline saltof Compound 1, or crystalline form thereof, may be adjusted over thecourse of the treatment, based on the judgment of the administeringphysician.

The choline salt of Compound 1, or crystalline form thereof may beadministered to a subject (e.g., a human) in an effective amount. Incertain embodiments, the choline salt of Compound 1, or crystalline formthereof is administered once daily.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, may be administered to a subject (e.g., a human) in antherapeutically effective amount. In some embodiments, the choline saltof Compound 1, or crystalline form thereof, is administered once daily.In some embodiments, the choline salt of Compound 1, or crystalline formthereof, is administered monthly. In some embodiments, the choline saltof Compound 1, or crystalline form thereof, is administered every threemonths. In some embodiments, the choline salt of Compound 1, orcrystalline form thereof, is administered every four months. In someembodiments, the choline salt of Compound 1, or crystalline formthereof, is administered every six months.

A choline salt of Compound 1, or crystalline form thereof, disclosedherein may be administered in a dosage amount that is effective. Forexample, the dosage amount can be from 1 mg to 1000 mg of compound. Incertain embodiments, the dosage amount is about 1, 10, 20, 30, 40, 50,60, 70, 80, 90, 95, 100, 105, 110, 120, 130, 140, or 150 mg of compound.In certain embodiments the dosage amount is about 100, 150, 200, 250,300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or1000 mg.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, is administered in a once daily dose. In some embodiments, thecholine salt of Compound 1, or crystalline form thereof, is administeredin a once daily dose of about 1 mg.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, is administered monthly. In some embodiments, the choline saltof Compound 1, or crystalline form thereof, is administered monthly at adose of about 100 mg.

In some embodiments, the choline salt of Compound 1, or crystalline formthereof, is administered every 6 months. In some embodiments, thecholine salt of Compound 1, or crystalline form thereof, is administeredevery 6 months at a dose of about 600 mg.

Combination Therapies

In certain embodiments, a method for treating or preventing an HIVinfection in a human having or at risk of having the infection isprovided, comprising administering to the human a therapeuticallyeffective amount of a choline salt of Compound 1, or crystalline formthereof, disclosed herein in combination with a therapeuticallyeffective amount of one or more (e.g., one, two, three, or four; or oneor two; or one to three; or one to four) additional therapeutic agents.In one embodiment, a method for treating an HIV infection in a humanhaving or at risk of having the infection is provided, comprisingadministering to the human a therapeutically effective amount of acholine salt of Compound 1, or crystalline form thereof, disclosedherein in combination with a therapeutically effective amount of one ormore (e.g., one, two, three, or four; or one or two; or one to three; orone to four) additional therapeutic agents.

In some embodiments, a method for treating or preventing an HIVinfection in a human having or at risk of having the infection isprovided, comprising administering to the human a therapeuticallyeffective amount of a choline salt of Compound 1, or crystalline formthereof, in combination with a therapeutically effective amount of oneor more (e.g., one, two, three, or four; or one or two; or one to three;or one to four) additional therapeutic agents. In one embodiment, amethod for treating an HIV infection in a human having or at risk ofhaving the infection is provided, comprising administering to the humana therapeutically effective amount of a choline salt of Compound 1, orcrystalline form thereof, in combination with a therapeuticallyeffective amount of one or more (e.g., one, two, three, or four; or oneor two; or one to three; or one to four) additional therapeutic agents.

In one embodiment, pharmaceutical compositions comprising a choline saltof Compound 1, or crystalline form thereof, in combination with one ormore (e.g., one, two, three, or four; or one or two; or one to three; orone to four) additional therapeutic agents, and a choline excipient areprovided.

In some embodiments, pharmaceutical compositions comprising a cholinesalt of Compound 1, or crystalline form thereof, in combination with oneor more (e.g., one, two, three, or four; or one or two; or one to three;or one to four) additional therapeutic agents, and a choline excipientare provided.

In certain embodiments, the present disclosure provides a method fortreating an HIV infection, comprising administering to a subject in needthereof a therapeutically effective amount of a choline salt of Compound1, or crystalline form thereof, in combination with a therapeuticallyeffective amount of one or more additional therapeutic agents which aresuitable for treating an HIV infection.

In certain embodiments, the present disclosure provides a method fortreating an HIV infection, comprising administering to a subject in needthereof a therapeutically effective amount of a choline salt of Compound1, or crystalline form thereof, in combination with a therapeuticallyeffective amount of one or more additional therapeutic agents which aresuitable for treating an HIV infection.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with one, two, three, four,or more additional therapeutic agents. In certain embodiments, a cholinesalt of Compound 1, or crystalline form thereof, disclosed herein iscombined with one additional therapeutic agent. In certain embodiments,a choline salt of Compound 1, or crystalline form thereof, disclosedherein is combined with two additional therapeutic agents. In otherembodiments, a choline salt of Compound 1, or crystalline form thereof,disclosed herein is combined with three additional therapeutic agents.In further embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with four additionaltherapeutic agents. The one, two, three, four, or more additionaltherapeutic agents can be different therapeutic agents selected from thesame class of therapeutic agents, and/or they can be selected fromdifferent classes of therapeutic agents.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with one, two, three, four, ormore additional therapeutic agents. In certain embodiments, a cholinesalt of Compound 1, or crystalline form thereof, disclosed herein iscombined with one additional therapeutic agent. In certain embodiments,a choline salt of Compound 1, or crystalline form thereof, disclosedherein is combined with two additional therapeutic agents. In otherembodiments, a choline salt of Compound 1, or crystalline form thereof,disclosed herein is combined with three additional therapeutic agents.In further embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with four additionaltherapeutic agents. The one, two, three, four, or more additionaltherapeutic agents can be different therapeutic agents selected from thesame class of therapeutic agents, and/or they can be selected fromdifferent classes of therapeutic agents.

Administration of HIV Combination Therapy

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is administered with one or moreadditional therapeutic agents. Co-administration of a choline salt ofCompound 1, or crystalline form thereof, disclosed herein with one ormore additional therapeutic agents generally refers to simultaneous orsequential administration of a choline salt of Compound 1, orcrystalline form thereof, and one or more additional therapeutic agents,such that therapeutically effective amounts of the choline salt ofCompound 1, or crystalline form thereof, and the one or more additionaltherapeutic agents are both present in the body of the subject. Whenadministered sequentially, the combination may be administered in two ormore administrations.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is administered with one or more additionaltherapeutic agents. Co-administration of a choline salt of Compound 1,or crystalline form thereof, disclosed herein with one or moreadditional therapeutic agents generally refers to simultaneous orsequential administration of the choline salt of Compound 1, orcrystalline form thereof, and one or more additional therapeutic agents,such that therapeutically effective amounts of the a choline salt ofCompound 1, or crystalline form thereof, and the one or more additionaltherapeutic agents are both present in the body of the subject. Whenadministered sequentially, the combination may be administered in two ormore administrations.

Co-administration includes administration of unit dosages of the cholinesalt of Compound 1, or crystalline form thereof, before or afteradministration of unit dosages of one or more additional therapeuticagents. For example, the choline salt of Compound 1, or crystalline formthereof, may be administered within seconds, minutes, or hours of theadministration of the one or more additional therapeutic agents. In someembodiments, a unit dose of a choline salt of Compound 1, or crystallineform thereof, is administered first, followed within seconds or minutesby administration of a unit dose of one or more additional therapeuticagents. Alternatively, a unit dose of one or more additional therapeuticagents is administered first, followed by administration of a unit doseof a choline salt of Compound 1, or crystalline form thereof, withinseconds or minutes. In other embodiments, a unit dose of a choline saltof Compound 1, or crystalline form thereof, is administered first,followed, after a period of hours (e.g., 1-12 hours), by administrationof a unit dose of one or more additional therapeutic agents. In yetother embodiments, a unit dose of one or more additional therapeuticagents is administered first, followed, after a period of hours (e.g.,1-12 hours), by administration of a unit dose of a choline salt ofCompound 1, or crystalline form thereof.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with one or more additionaltherapeutic agents in a unitary dosage form for simultaneousadministration to a subject. In certain embodiments, such a unitarydosage form can be administered by any route appropriate to thecondition to be treated. Suitable routes include oral, rectal, nasal,topical (including buccal and sublingual), transdermal, vaginal andparenteral (including subcutaneous, intramuscular, intravenous,intradermal, intrathecal and epidural), and the like. In certainembodiments, the compounds disclosed can be dosed parenterally. Incertain embodiments, the unitary dosage form can be dosed intravenous,subcutaneous, or intramuscular. In certain embodiments, the unitarydosage form is orally bioavailable and can be dosed orally. In certainembodiments, the unitary dosage form can be a solid dosage form for oraladministration.

The choline salt of Compound 1, or crystalline form thereof, disclosedherein in combination with one or more additional therapeutic agents canbe administered by any route appropriate to the condition to be treated.Suitable routes include oral, rectal, nasal, topical (including buccaland sublingual), transdermal, vaginal and parenteral (includingsubcutaneous, intramuscular, intravenous, intradermal, intrathecal andepidural), and the like. In certain embodiments, the choline salt ofCompound 1, or crystalline form thereof, disclosed herein can be dosedparenterally. In certain embodiments, the choline salt of Compound 1, orcrystalline form thereof, disclosed herein can be dosed intravenous,subcutaneous, or intramuscular. In certain embodiments, the choline saltof Compound 1, or crystalline form thereof, disclosed herein are orallybioavailable and can be dosed orally.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is formulated as a tablet, which mayoptionally contain one or more other compounds useful for treating HIV.In certain embodiments, the tablet can one or more other compoundsuseful for treating HIV, such as HIV protease inhibitors, HIVnon-nucleoside or non-nucleotide inhibitors of reverse transcriptase,HIV nucleoside or nucleotide inhibitors of reverse transcriptase, HIVintegrase inhibitors, HIV non-catalytic site (or allosteric) integraseinhibitors, pharmacokinetic enhancers, and combinations thereof.

In certain embodiments, a pharmaceutically acceptable salt of Compound1, or a cocrystal or crystalline form thereof, disclosed herein isformulated as a tablet, which may optionally contain one or more othercompounds useful for treating HIV. In certain embodiments, the tabletcan one or more other compounds useful for treating HIV, such as HIVnucleoside reverse transcriptase translocation inhibitors, HIV proteaseinhibitors, HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase, HIV nucleoside or nucleotide inhibitors of reversetranscriptase, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, pharmacokinetic enhancers, andcombinations thereof.

In certain embodiments, such tablets are suitable for once daily dosing.

HIV Combination Therapy

In some embodiments, a pharmaceutically acceptable salt of Compound 1,or a cocrystal or crystalline form thereof, disclosed herein isadministered with at least one additional therapeutic agent.

In the above embodiments, the additional therapeutic agent may be ananti-HIV agent selected from the group consisting of combination drugsfor treating HIV, other drugs for treating HIV, HIV protease inhibitors,HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase, HIV nucleoside or nucleotide inhibitors of reversetranscriptase, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, HIV entry inhibitors, HIV maturationinhibitors, latency reversing agents, compounds that target the HIVcapsid, immune-based therapies, phosphatidylinositol 3-kinase (PI3K)inhibitors, HIV antibodies, bispecific antibodies and “antibody-like”therapeutic proteins, HIV p17 matrix protein inhibitors, IL-13antagonists, peptidyl-prolyl cis-trans isomerase A modulators, proteindisulfide isomerase inhibitors, complement C5a receptor antagonists, DNAmethyltransferase inhibitor, HIV vif gene modulators, Vif dimerizationantagonists, HIV-1 viral infectivity factor inhibitors, TAT proteininhibitors, HIV-1 Nef modulators, Hck tyrosine kinase modulators, mixedlineage kinase-3 (MLK-3) inhibitors, HIV-1 splicing inhibitors, Revprotein inhibitors, integrin antagonists, nucleoprotein inhibitors,splicing factor modulators, COMM domain containing protein 1 modulators,HIV ribonuclease H inhibitors, retrocyclin modulators, CDK-9 inhibitors,dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAG proteininhibitors, HIV POL protein inhibitors, Complement Factor H modulators,ubiquitin ligase inhibitors, deoxycytidine kinase inhibitors, cyclindependent kinase inhibitors, proprotein convertase PC9 stimulators, ATPdependent RNA helicase DDX3X inhibitors, reverse transcriptase primingcomplex inhibitors, G6PD and NADH-oxidase inhibitors, pharmacokineticenhancers, HIV gene therapy, HIV vaccines, and combinations thereof.

In some embodiments, the additional therapeutic agent is selected fromthe group consisting of combination drugs for HIV, other drugs fortreating HIV, HIV protease inhibitors, HIV reverse transcriptaseinhibitors, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIVmaturation inhibitors, latency reversing agents, capsid inhibitors,immune-based therapies, PI3K inhibitors, HIV antibodies, and bispecificantibodies, and “antibody-like” therapeutic proteins, and combinationsthereof.

In some embodiments, the additional therapeutic agent is selected fromthe group consisting of combination drugs for HIV, other drugs fortreating HIV, HIV nucleoside reverse transcriptase translocationinhibitors, HIV protease inhibitors, HIV reverse transcriptaseinhibitors, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIVmaturation inhibitors, latency reversing agents, capsid inhibitors,immune-based therapies, PI3K inhibitors, HIV antibodies, and bispecificantibodies, and “antibody-like” therapeutic proteins, and combinationsthereof. In some embodiments, the additional therapeutic agent isselected from immunomodulators, immunotherapeutic agents, antibody-drugconjugates, gene modifiers, gene editors (such as CRISPR/Cas9, zincfinger nucleases, homing nucleases, synthetic nucleases, TALENs), andcell therapies such as chimeric antigen receptor T-cell, CAR-T (e.g.,YESCARTA® (axicabtagene ciloleucel)), and engineered T cell receptors,TCR-T.

HIV Combination Drugs

Examples of combination drugs include ATRIPLA® (efavirenz, tenofovirdisoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®;rilpivirine, tenofovir disoproxil fumarate, and emtricitabine);STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, andemtricitabine); TRUVADA® (tenofovir disoproxil fumarate andemtricitabine; TDF+FTC); DESCOVY® (tenofovir alafenamide andemtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, andrilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine,cobicistat, and elvitegravir); darunavir, tenofovir alafenamidehemifumarate, emtricitabine, and cobicistat; efavirenz, lamivudine, andtenofovir disoproxil fumarate; lamivudine and tenofovir disoproxilfumarate; tenofovir and lamivudine; tenofovir alafenamide andemtricitabine; tenofovir alafenamide hemifumarate and emtricitabine;tenofovir alafenamide hemifumarate, emtricitabine, and rilpivirine;tenofovir alafenamide hemifumarate, emtricitabine, cobicistat, andelvitegravir; COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM®(LIVEXA®; abacavir sulfate and lamivudine; ABC+3TC); KALETRA® (ALUVIA®;lopinavir and ritonavir); TRIUMEQ® (dolutegravir, abacavir, andlamivudine); TRIZIVIR® (abacavir sulfate, zidovudine, and lamivudine;ABC+AZT+3TC); atazanavir and cobicistat; atazanavir sulfate andcobicistat; atazanavir sulfate and ritonavir; darunavir and cobicistat;dolutegravir and rilpivirine; dolutegravir and rilpivirinehydrochloride; cabotegravir and rilpivirine; cabotegravir andrilpivirine hydrochloride; dolutegravir, abacavir sulfate, andlamivudine; lamivudine, nevirapine, and zidovudine; raltegravir andlamivudine; doravirine, lamivudine, and tenofovir disoproxil fumarate;doravirine, lamivudine, and tenofovir disoproxil;dolutegravir+lamivudine; lamivudine+abacavir+zidovudine;lamivudine+abacavir; lamivudine+tenofovir disoproxil fumarate;lamivudine+zidovudine+nevirapine; lopinavir+ritonavir;lopinavir+ritonavir+abacavir+lamivudine;lopinavir+ritonavir+zidovudine+lamivudine; tenofovir+lamivudine; andtenofovir disoproxil fumarate+emtricitabine+rilpivirine hydrochloride;lopinavir, ritonavir, zidovudine and lamivudine; Vacc-4x and romidepsin;and APH-0812.

Other HIV Drugs

Examples of other drugs for treating HIV include acemannan, alisporivir,BanLec, deferiprone, Gamimune, metenkefalin, naltrexone, Prolastin, REP9, RPI-MN, VSSP, Hlviral, SB-728-T, 1,5-dicaffeoylquinic acid,rHIV7-shl-TAR-CCRSRZ, AAV-eCD4-Ig gene therapy, MazF gene therapy,BlockAide, ABX-464, AG-1105, APH-0812, BIT-225, CYT-107, HGTV-43,HPH-116, HS-10234, IMO-3100, IND-02, MK-1376, MK-8507, MK-8591, NOV-205,PA-1050040 (PA-040), PGN-007, SCY-635, SB-9200, SCB-719, TR-452,TEV-90110, TEV-90112, TEV-90111, TEV-90113, RN-18, Immuglo, and VIR-576.

HIV Nucleoside Reverse Transcriptase Translocation Inhibitors

Examples of HIV nucleoside reverse transcriptase translocationinhibitors (“NRTTIs”) include 4′-Ethynyl-2-fluoro-2′-deoxyadenosinetriphosphate (also known as MK-8591 and EFdA).

HIV Protease Inhibitors

Examples of HIV protease inhibitors include amprenavir, atazanavir,brecanavir, darunavir, fosamprenavir, fosamprenavir calcium, indinavir,indinavir sulfate, lopinavir, nelfinavir, nelfinavir mesylate,ritonavir, saquinavir, saquinavir mesylate, tipranavir, DG-17, TMB-657(PPL-100), T-169, BL-008, and TMC-310911.

HIV Reverse Transcriptase Inhibitors

Examples of HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase include dapivirine, delavirdine, delavirdine mesylate,doravirine, efavirenz, etravirine, lentinan, nevirapine, rilpivirine,AIC-292, KM-023, and VM-1500. Further examples of non-nucleoside reversetranscriptase inhibitors are disclosed in U.S. Patent Publication No.US2016/0250215.

Examples of HIV nucleoside or nucleotide inhibitors of reversetranscriptase include adefovir, adefovir dipivoxil, azvudine,emtricitabine, tenofovir, tenofovir alafenamide, tenofovir alafenamidefumarate, tenofovir alafenamide hemifumarate, tenofovir disoproxil,tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, VIDEX®and VIDEX EC® (didanosine, ddl), abacavir, abacavir sulfate, alovudine,apricitabine, censavudine, didanosine, elvucitabine, festinavir,fosalvudine tidoxil, CMX-157, dapivirine, doravirine, etravirine,OCR-5753, tenofovir disoproxil orotate, fozivudine tidoxil, lamivudine,phosphazid, stavudine, zalcitabine, zidovudine, GS-9131, GS-9148, andKP-1461.

In some embodiments, examples of HIV nucleoside or nucleotide inhibitorsof reverse transcriptase include adefovir, adefovir dipivoxil, azvudine,emtricitabine, tenofovir, tenofovir alafenamide, tenofovir alafenamidefumarate, tenofovir alafenamide hemifumarate, tenofovir disoproxil,tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, VIDEX®and VIDEX EC® (didanosine, ddl), abacavir, abacavir sulfate, alovudine,apricitabine, censavudine, didanosine, elvucitabine, festinavir,fosalvudine tidoxil, CMX-157, dapivirine, doravirine, etravirine,OCR-5753, tenofovir disoproxil orotate, fozivudine tidoxil, lamivudine,phosphazid, stavudine, zalcitabine, zidovudine, GS-9131, GS-9148,KP-1461, and 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA).

HIV Integrase Inhibitors

Examples of HIV integrase inhibitors include elvitegravir, curcumin,derivatives of curcumin, chicoric acid, derivatives of chicoric acid,3,5-dicaffeoylquinic acid, derivatives of 3,5-dicaffeoylquinic acid,aurintricarboxylic acid, derivatives of aurintricarboxylic acid, caffeicacid phenethyl ester, derivatives of caffeic acid phenethyl ester,tyrphostin, derivatives of tyrphostin, quercetin, derivatives ofquercetin, raltegravir, dolutegravir, JTK-351, bictegravir, AVX-15567,diketo quinolin-4-1 derivatives, integrase-LEDGF inhibitor, ledgins,M-522, M-532, NSC-310217, NSC-371056, NSC-48240, NSC-642710, NSC-699171,NSC-699172, NSC-699173, NSC-699174, stilbenedisulfonic acid, T-169 andcabotegravir.

Examples of HIV non-catalytic site, or allosteric, integrase inhibitors(NCINI) include CX-05045, CX-05168, and CX-14442.

HIV Entry Inhibitors

Examples of HIV entry (fusion) inhibitors include cenicriviroc, CCR5inhibitors, gp41 inhibitors, CD4 attachment inhibitors, gp120inhibitors, and CXCR4 inhibitors.

Examples of CCR5 inhibitors include aplaviroc, vicriviroc, maraviroc,cenicriviroc, PRO-140, adaptavir (RAP-101), nifeviroc (TD-0232),anti-GP120/CD4 or CCR5 bispecific antibodies, B-07, MB-66, polypeptideC25P, TD-0680, and vMIP (Haimipu).

Examples of gp41 inhibitors include albuvirtide, enfuvirtide,BMS-986197, enfuvirtide biobetter, enfuvirtide biosimilar, HIV-1 fusioninhibitors (P26-Bapc), ITV-1, ITV-2, ITV-3, ITV-4, PIE-12 trimer andsifuvirtide.

Examples of CD4 attachment inhibitors include ibalizumab and CADAanalogs

Examples of gp120 inhibitors include Radha-108 (receptol) 3B3-PE38,BanLec, bentonite-based nanomedicine, fostemsavir tromethamine,IQP-0831, and BMS-663068

Examples of CXCR4 inhibitors include plerixafor, ALT-1188, N15 peptide,and vMIP (Haimipu).

HIV Maturation Inhibitors

Examples of HIV maturation inhibitors include BMS-955176 andGSK-2838232.

Latency Reversing Agents

Examples of latency reversing agents include histone deacetylase (HDAC)inhibitors, proteasome inhibitors such as velcade, protein kinase C(PKC) activators, BET-bromodomain 4 (BRD4) inhibitors, ionomycin, PMA,SAHA (suberanilohydroxamic acid, or suberoyl, anilide, and hydroxamicacid), IL-15, JQ1, disulfram, amphotericin B, and ubiquitin inhibitorssuch as largazole analogs, and GSK-343.

Examples of HDAC inhibitors include romidepsin, vorinostat, andpanobinostat.

Examples of PKC activators include indolactam, prostratin, ingenol B,and DAG-lactones.

Capsid Inhibitors

Examples of capsid inhibitors include capsid polymerization inhibitorsor capsid disrupting compounds, HIV nucleocapsid p7 (NCp7) inhibitorssuch as azodicarbonamide, HIV p24 capsid protein inhibitors, AVI-621,AVI-101, AVI-201, AVI-301, and AVI-CAN1-15 series.

Immune-Based Therapies

Examples of immune-based therapies include toll-like receptorsmodulators such as tlr1, tlr2, tlr3, tlr4, tlr5, tlr6, tlr7, tlr8, tlr9,tlr10, tlr11, tlr12, and tlr13; programmed cell death protein 1 (Pd-1)modulators; programmed death-ligand 1 (Pd-L1) modulators; IL-15agonists; DermaVir; interleukin-7; plaquenil (hydroxychloroquine);proleukin (aldesleukin, IL-2); interferon alfa; interferon alfa-2b;interferon alfa-n3; pegylated interferon alfa; interferon gamma;hydroxyurea; mycophenolate mofetil (MPA) and its ester derivativemycophenolate mofetil (MMF); ribavirin; rintatolimod, polymerpolyethyleneimine (PEI); gepon; rintatolimod; IL-12; WF-10; VGV-1;MOR-22; BMS-936559; CYT-107, interleukin-15/Fc fusion protein,normferon, peginterferon alfa-2a, peginterferon alfa-2b, recombinantinterleukin-15, RPI-MN, GS-9620, and IR-103.

Phosphatidylinositol 3-Kinase (PI3K) Inhibitors

Examples of PI3K inhibitors include idelalisib, alpelisib, buparlisib,CAI orotate, copanlisib, duvelisib, gedatolisib, neratinib, panulisib,perifosine, pictilisib, pilaralisib, puquitinib mesylate, rigosertib,rigosertib sodium, sonolisib, taselisib, AMG-319, AZD-8186, BAY-1082439,CLR-1401, CLR-457, CUDC-907, DS-7423, EN-3342, GSK-2126458, GSK-2269577,GSK-2636771, INCB-040093, LY-3023414, MLN-1117, PQR-309, RG-7666,RP-6530, RV-1729, SAR-245409, SAR-260301, SF-1126, TGR-1202, UCB-5857,VS-5584, XL-765, and ZSTK-474.

HIV Antibodies, Bispecific Antibodies, and “Antibody-Like” TherapeuticProteins

Examples of HIV antibodies, bispecific antibodies, and “antibody-like”therapeutic proteins include DARTs®, DUOBODIES®, BITES®, XmAbs®,TandAbs®, Fab derivatives, bnABs (broadly neutralizing HIV-1antibodies), BMS-936559, TMB-360, and those targeting HIV gp120 or gp41,antibody-Recruiting Molecules targeting HIV, anti-CD63 monoclonalantibodies, anti-GB virus C antibodies, anti-GP120/CD4, CCR5 bispecificantibodies, anti-nef single domain antibodies, anti-Rev antibody,camelid derived anti-CD18 antibodies, camelid-derived anti-ICAM-1antibodies, DCVax-001, gp140 targeted antibodies, gp41-based HIVtherapeutic antibodies, human recombinant mAbs (PGT-121), ibalizumab,Immuglo, MB-66

Examples of those targeting HIV in such a manner include bavituximab,UB-421, C2F5, C2G12, C4E10, C2F5+C2G12+C4E10, 3-BNC-117, PGT145, PGT121,MDX010 (ipilimumab), VRC01, A32, 7B2, 10E8, VRC-07-523,VRC-HIVMAB080-00-AB, MGD-014 and VRC07.

Pharmacokinetic Enhancers

Examples of pharmacokinetic enhancers include cobicistat and ritonavir.

Additional Therapeutic Agents

Examples of additional therapeutic agents include the compoundsdisclosed in WO 2004/096286 (Gilead Sciences), WO 2006/015261 (GileadSciences), WO 2006/110157 (Gilead Sciences), WO 2012/003497 (GileadSciences), WO 2012/003498 (Gilead Sciences), WO 2012/145728 (GileadSciences), WO 2013/006738 (Gilead Sciences), WO 2013/159064 (GileadSciences), WO 2014/100323 (Gilead Sciences), US 2013/0165489 (Universityof Pennsylvania), US 2014/0221378 (Japan Tobacco), US 2014/0221380(Japan Tobacco), WO 2009/062285 (Boehringer Ingelheim), WO 2010/130034(Boehringer Ingelheim), WO 2013/006792 (Pharma Resources), US20140221356 (Gilead Sciences), US 20100143301 (Gilead Sciences) and WO2013/091096 (Boehringer Ingelheim).

HIV Vaccines

Examples of HIV vaccines include peptide vaccines, recombinant subunitprotein vaccines, live vector vaccines, DNA vaccines, CD4-derivedpeptide vaccines, vaccine combinations, rgp120 (AIDSVAX), ALVAC HIV(vCP1521)/AIDSVAX B/E (gp120) (RV144), monomeric gp120 HIV-1 subtype Cvaccine, Remune, ITV-1, Contre Vir, Ad5-ENVA-48, DCVax-001 (CDX-2401),Vacc-4x, Vacc-05, VAC-3S, multiclade DNA recombinant adenovirus-5(rAd5), Pennvax-G, Pennvax-GP, HIV-TriMix-mRNA vaccine, HIV-LAMP-vax,Ad35, Ad35-GRIN, NAcGM3/VSSP ISA-51, poly-ICLC adjuvanted vaccines,TatImmune, GTU-multiHIV (FIT-06), gp140[delta]V2.TV1+MF-59, rVSVIN HIV-1gag vaccine, SeV-Gag vaccine, AT-20, DNK-4, ad35-Grin/ENV, TBC-M4,HIVAX, HIVAX-2, NYVAC-HIV-PT1, NYVAC-HIV-PT4, DNA-HIV-PT123,rAAV1-PG9DP, GOVX-B11, GOVX-B21, TVI-HIV-1, Ad-4 (Ad4-env CladeC+Ad4-mGag), EN41-UGR7C, EN41-FPA2, PreVaxTat, AE-H, MYM-V101,CombiHIVvac, ADVAX, MYM-V201, MVA-CMDR, DNA-Ad5 gag/pol/nef/nev(HVTN505), MVATG-17401, ETV-01, CDX-1401, rcAD26.MOS1.HIV-Env,Ad26.Mod.HIV vaccine, AGS-004, AVX-101, AVX-201, PEP-6409, SAV-001,ThV-01, TL-01, TUTI-16, VGX-3300, IHV-001, and virus-like particlevaccines such as pseudovirion vaccine, CombiVICHvac, LFn-p24 B/C fusionvaccine, GTU-based DNA vaccine, HIV gag/pol/nef/env DNA vaccine,anti-TAT HIV vaccine, conjugate polypeptides vaccine, dendritic-cellvaccines, gag-based DNA vaccine, GI-2010, gp41 HIV-1 vaccine, HIVvaccine (PIKA adjuvant), I i-key/MHC class II epitope hybrid peptidevaccines, ITV-2, ITV-3, ITV-4, LIPO-5, multiclade Env vaccine, MVAvaccine, Pennvax-GP, pp71-deficient HCMV vector HIV gag vaccine,recombinant peptide vaccine (HIV infection), NCI, rgp160 HIV vaccine,RNActive HIV vaccine, SCB-703, Tat Oyi vaccine, TBC-M4, therapeutic HIVvaccine, UBI HIV gp120, Vacc-4x+romidepsin, variant gp120 polypeptidevaccine, rAd5 gag-pol env A/B/C vaccine.

HIV Combination Therapy

In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with one, two, three, four ormore additional therapeutic agents selected from ATRIPLA® (efavirenz,tenofovir disoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®;rilpivirine, tenofovir disoproxil fumarate, and emtricitabine);STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, andemtricitabine); TRUVADA® (tenofovir disoproxil fumarate andemtricitabine; TDF+FTC); DESCOVY® (tenofovir alafenamide andemtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, andrilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine,cobicistat, and elvitegravir); adefovir; adefovir dipivoxil; cobicistat;emtricitabine; tenofovir; tenofovir disoproxil; tenofovir disoproxilfumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate;TRIUMEQ® (dolutegravir, abacavir, and lamivudine); dolutegravir,abacavir sulfate, and lamivudine; raltegravir; raltegravir andlamivudine; maraviroc; enfuvirtide; ALUVIA® (KALETRA®; lopinavir andritonavir); COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM®(LIVEXA®; abacavir sulfate and lamivudine; ABC+3TC); TRIZIVIR® (abacavirsulfate, zidovudine, and lamivudine; ABC+AZT+3TC); rilpivirine;rilpivirine hydrochloride; atazanavir sulfate and cobicistat; atazanavirand cobicistat; darunavir and cobicistat; atazanavir; atazanavirsulfate; dolutegravir; elvitegravir; ritonavir; atazanavir sulfate andritonavir; darunavir; lamivudine; prolastin; fosamprenavir;fosamprenavir calcium efavirenz; etravirine; nelfinavir; nelfinavirmesylate; interferon; didanosine; stavudine; indinavir; indinavirsulfate; tenofovir and lamivudine; zidovudine; nevirapine; saquinavir;saquinavir mesylate; aldesleukin; zalcitabine; tipranavir; amprenavir;delavirdine; delavirdine mesylate; Radha-108 (receptol); lamivudine andtenofovir disoproxil fumarate; efavirenz, lamivudine, and tenofovirdisoproxil fumarate; phosphazid; lamivudine, nevirapine, and zidovudine;abacavir; and abacavir sulfate.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with one, two, three, four or moreadditional therapeutic agents selected from ATRIPLA® (efavirenz,tenofovir disoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®;rilpivirine, tenofovir disoproxil fumarate, and emtricitabine);STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, andemtricitabine); TRUVADA® (tenofovir disoproxil fumarate andemtricitabine; TDF+FTC); DESCOVY® (tenofovir alafenamide andemtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, andrilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine,cobicistat, and elvitegravir); adefovir; adefovir dipivoxil; cobicistat;emtricitabine; tenofovir; tenofovir disoproxil; tenofovir disoproxilfumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate;TRIUMEQ® (dolutegravir, abacavir, and lamivudine); dolutegravir,abacavir sulfate, and lamivudine; raltegravir; raltegravir andlamivudine; maraviroc; enfuvirtide; ALUVIA® (KALETRA®; lopinavir andritonavir); COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM®(LIVEXA®; abacavir sulfate and lamivudine; ABC+3TC); TRIZIVIR® (abacavirsulfate, zidovudine, and lamivudine; ABC+AZT+3TC); rilpivirine;rilpivirine hydrochloride; atazanavir sulfate and cobicistat; atazanavirand cobicistat; darunavir and cobicistat; atazanavir; atazanavirsulfate; dolutegravir; elvitegravir; ritonavir; atazanavir sulfate andritonavir; darunavir; lamivudine; prolastin; fosamprenavir;fosamprenavir calcium efavirenz; etravirine; nelfinavir; nelfinavirmesylate; interferon; didanosine; stavudine; indinavir; indinavirsulfate; tenofovir and lamivudine; zidovudine; nevirapine; saquinavir;saquinavir mesylate; aldesleukin; zalcitabine; tipranavir; amprenavir;delavirdine; delavirdine mesylate; Radha-108 (receptol); lamivudine andtenofovir disoproxil fumarate; efavirenz, lamivudine, and tenofovirdisoproxil fumarate; phosphazid; lamivudine, nevirapine, and zidovudine;abacavir; abacavir sulfate; 4′-ethynyl-2-fluoro-2′-deoxyadenosine(EFdA); and Bictegravir, or a pharmaceutically acceptable salt thereof.

It will be appreciated by one of skill in the art that the additionaltherapeutic agents listed above may be included in more than one of theclasses listed above. The particular classes are not intended to limitthe functionality of those compounds listed in those classes.

In a specific embodiment a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with one or two HIVnucleoside or nucleotide inhibitors of reverse transcriptase. In aspecific embodiment, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with an HIV nucleoside ornucleotide inhibitor of reverse transcriptase and an HIV non-nucleosideinhibitor of reverse transcriptase. In another specific embodiment, acholine salt of Compound 1, or crystalline form thereof, disclosedherein is combined with an HIV nucleoside or nucleotide inhibitor ofreverse transcriptase, and an HIV protease inhibiting compound. In anadditional embodiment, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with an HIV nucleoside ornucleotide inhibitor of reverse transcriptase, an HIV non-nucleosideinhibitor of reverse transcriptase, and a pharmacokinetic enhancer. Incertain embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with at least one HIV nucleosideinhibitor of reverse transcriptase, an integrase inhibitor, and apharmacokinetic enhancer.

In another embodiment, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with two HIV nucleoside ornucleotide inhibitors of reverse transcriptase.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with one or two HIV nucleoside ornucleotide inhibitors of reverse transcriptase. In a specificembodiment, a choline salt of Compound 1, or crystalline form thereof,disclosed herein is combined with an HIV nucleoside or nucleotideinhibitor of reverse transcriptase and an HIV non-nucleoside inhibitorof reverse transcriptase. In another specific embodiment, a choline saltof Compound 1, or crystalline form thereof, disclosed herein is combinedwith an HIV nucleoside or nucleotide inhibitor of reverse transcriptase,and an HIV protease inhibiting compound. In an additional embodiment, acholine salt of Compound 1, or crystalline form thereof, disclosedherein is combined with an HIV nucleoside or nucleotide inhibitor ofreverse transcriptase, an HIV non-nucleoside inhibitor of reversetranscriptase, and a pharmacokinetic enhancer. In certain embodiments, acholine salt of Compound 1, or crystalline form thereof, disclosedherein is combined with at least one HIV nucleoside inhibitor of reversetranscriptase, an integrase inhibitor, and a pharmacokinetic enhancer.In another embodiment, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with two HIV nucleoside ornucleotide inhibitors of reverse transcriptase.

In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with abacavir sulfate,tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate,tenofovir disoproxil hemifumarate, tenofovir alafenamide, tenofoviralafenamide fumarate or tenofovir alafenamide hemifumarate.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with abacavir sulfate, tenofovir,tenofovir disoproxil, tenofovir disoproxil fumarate, tenofovirdisoproxil hemifumarate, tenofovir alafenamide, tenofovir alafenamidefumarate, tenofovir alafenamide hemifumarate, bictegravir (or a cholinesalt thereof), or 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA).

In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with tenofovir, tenofovirdisoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide,tenofovir alafenamide fumarate or tenofovir alafenamide hemifumarate.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with tenofovir, tenofovirdisoproxil, tenofovir disoproxil fumarate, tenofovir alafenamide,tenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate,bictegravir (or a pharmaceutically acceptable salt thereof), or4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA).

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with a first additionaltherapeutic agent selected from the group consisting of abacavirsulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate,tenofovir alafenamide, tenofovir alafenamide fumarate and tenofoviralafenamide hemifumarate, and a second additional therapeutic agentselected from the group consisting of emtricitabine and lamivudine.

In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with a first additionaltherapeutic agent selected from the group consisting of tenofovir,tenofovir disoproxil, tenofovir disoproxil fumarate, tenofoviralafenamide, and tenofovir alafenamide hemifumarate, and a secondadditional therapeutic agent, wherein the second additional therapeuticagent is emtricitabine. In a particular embodiment, a choline salt ofCompound 1, or crystalline form thereof, disclosed herein is combinedwith a first additional therapeutic agent selected from the groupconsisting of tenofovir alafenamide fumarate, tenofovir alafenamide, andtenofovir alafenamide hemifumarate, and a second additional therapeuticagent, wherein the second additional therapeutic agent is emtricitabine.In a particular embodiment, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with a first additionaltherapeutic agent selected from the group consisting of tenofovirdisoproxil fumarate, tenofovir disoproxil, and tenofovir disoproxilhemifumarate, and a second additional therapeutic agent, wherein thesecond additional therapeutic agent is emtricitabine. In someembodiments, the choline salt of Compound 1, or crystalline formthereof, and the first and second additional therapeutic agents asdisclosed above are administered simultaneously. Optionally, the cholinesalt of Compound 1, or crystalline form thereof, and the first andsecond additional therapeutic agents as disclosed above are combined ina unitary dosage form for simultaneous administration to a subject. Inother embodiments, the choline salt of Compound 1, or crystalline formthereof, and the first and second additional therapeutic agents asdisclosed above are administered sequentially.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with bictegravir or apharmaceutically acceptable salt thereof.

In some embodiments, a pharmaceutically acceptable salt of Compound 1,or a cocrystal or crystalline form thereof, disclosed herein is combinedwith 4′-ethynyl-2-fluoro-2′-deoxyadenosine (EFdA).

A choline salt of Compound 1, or crystalline form thereof, disclosedherein may be combined with one or more additional therapeutic agents inany dosage amount of the choline salt of Compound 1, or crystalline formthereof (e.g., from 1 mg to 1000 mg of the salt or crystalline form).

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein may be combined with one or more additionaltherapeutic agents in any dosage amount of the choline salt of Compound1, or crystalline form thereof (e.g., from 1 mg to 1000 mg of the saltor crystalline form).

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein is combined with 5-30 mg tenofoviralafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofoviralafenamide, and 200 mg emtricitabine. In certain embodiments, a cholinesalt of Compound 1, or crystalline form thereof, disclosed herein iscombined with 5-10, 5-15, 5-20, 5-25, 25-30, 20-30, 15-30, or 10-30 mgtenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, ortenofovir alafenamide, and 200 mg emtricitabine. In certain embodiments,a choline salt of Compound 1, or crystalline form thereof, disclosedherein is combined with 10 mg tenofovir alafenamide fumarate, tenofoviralafenamide hemifumarate, or tenofovir alafenamide, and 200 mgemtricitabine. In certain embodiments, a choline salt of Compound 1, orcrystalline form thereof, disclosed herein is combined with 25 mgtenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, ortenofovir alafenamide, and 200 mg emtricitabine. A choline salt ofCompound 1, or crystalline form thereof, disclosed herein may becombined with the agents provided herein in any dosage amount of thesalt or crystalline form (e.g., from 1 mg to 1000 mg of the salt orcrystalline form) the same as if each combination of dosages werespecifically and individually listed.

In certain embodiments, a choline salt of Compound 1, or crystallineform thereof, disclosed herein, is combined with 200-400 mg tenofovirdisoproxil fumarate, tenofovir disoproxil hemifumarate, or tenofovirdisoproxil, and 200 mg emtricitabine. In certain embodiments, a cholinesalt of Compound 1, or crystalline form thereof, disclosed herein iscombined with 200-250, 200-300, 200-350, 250-350, 250-400, 350-400,300-400, or 250-400 mg tenofovir disoproxil fumarate, tenofovirdisoproxil hemifumarate, or tenofovir disoproxil, and 200 mgemtricitabine. In certain embodiments, a choline salt of Compound 1, orcrystalline form thereof, disclosed herein is combined with 300 mgtenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, ortenofovir disoproxil, and 200 mg emtricitabine. A choline salt ofCompound 1, or crystalline form thereof, disclosed herein may becombined with the agents provided herein in any dosage amount of thesalt or crystalline form (e.g., from 1 mg to 1000 mg of the salt orcrystalline form) the same as if each combination of dosages werespecifically and individually listed.

In some embodiments, a choline salt of Compound 1, or crystalline formthereof, disclosed herein is combined with 20-80 mg of bictegravir or acholine salt thereof. A choline salt of Compound 1, or crystalline formthereof, disclosed herein may be combined with the agents providedherein in any dosage amount of the salt or crystalline form (e.g., from1 mg to 1000 mg of the salt or crystalline form) the same as if eachcombination of dosages were specifically and individually listed.

In one embodiment, kits comprising a choline salt of Compound 1, orcrystalline form thereof, disclosed herein in combination with one ormore (e.g., one, two, three, one or two, or one to three) additionaltherapeutic agents are provided.

In some embodiments, kits comprising a choline salt of Compound 1, orcrystalline form thereof, disclosed herein in combination with one ormore (e.g., one, two, three, one or two, or one to three) additionaltherapeutic agents are provided.

Pharmaceutical Compositions

Pharmaceutical compositions disclosed herein comprise a choline salt ofCompound 1, or crystalline form thereof, disclosed herein together withone or more pharmaceutically acceptable excipients and optionally othertherapeutic agents. Pharmaceutical compositions containing the activeingredient may be in any form suitable for the intended method ofadministration.

In some embodiments, pharmaceutical compositions disclosed hereincomprise a choline salt of Compound 1, or crystalline form thereof,disclosed herein together with one or more pharmaceutically acceptableexcipients and optionally other therapeutic agents. Pharmaceuticalcompositions containing the active ingredient may be in any formsuitable for the intended method of administration.

Pharmaceutical compositions comprising the choline salt of Compound 1,or crystalline form thereof, disclosed herein may be prepared withconventional carriers (e.g., inactive ingredient or excipient material)which may be selected in accord with ordinary practice. Tablets maycontain excipients including glidants, fillers, binders and the like.Aqueous compositions may be prepared in sterile form, and when intendedfor delivery by other than oral administration generally may beisotonic. All compositions may optionally contain excipients such asthose set forth in the Rowe et al, Handbook of PharmaceuticalExcipients, 5th edition, American Pharmacists Association, 1986.Excipients can include ascorbic acid and other antioxidants, chelatingagents such as EDTA, carbohydrates such as dextrin,hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid and thelike.

While it is possible for the active ingredient to be administered alone,it may be preferable to present the active ingredient as pharmaceuticalcompositions. The compositions, both for veterinary and for human use,comprise at least the choline salt of Compound 1, or crystalline formthereof, disclosed herein together with one or more acceptable carriersand optionally other therapeutic ingredients. In one embodiment, thepharmaceutical composition comprises a choline salt of Compound 1, orcrystalline form thereof, a pharmaceutically acceptable excipient, and atherapeutically effective amount of one or more (e.g., one, two, three,or four; or one or two; or one to three; or one to four) additionaltherapeutic agents as defined hereinbefore. In one embodiment, thepharmaceutical composition comprises a choline salt of Compound 1, orcrystalline form thereof, a pharmaceutically acceptable excipient, andone other therapeutic ingredient. The carrier(s) are “acceptable” in thesense of being compatible with the other ingredients of the compositionand physiologically innocuous to the recipient thereof.

The compositions include those suitable for various administrationroutes. The compositions may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Such methods include the step of bringing into association theactive ingredient with one or more inactive ingredients (e.g., acarrier, pharmaceutical excipient, etc.). The compositions may beprepared by uniformly and intimately bringing into association theactive ingredient with liquid carriers or finely divided solid carriersor both, and then, if necessary, shaping the product. Techniques andformulations generally are found in Remington: The Science and Practiceof Pharmacy, 21st Edition, Lippincott Wiliams and Wilkins, Philadelphia,Pa., 2006.

Compositions described herein that are suitable for oral administrationmay be presented as discrete units (a unit dosage form) including butnot limited to capsules, cachets or tablets each containing apredetermined amount of the active ingredient.

When used for oral use for example, tablets, troches, lozenges, aqueousor oil suspensions, dispersible powders or granules, emulsions, hard orsoft capsules, syrups or elixirs may be prepared. Compositions intendedfor oral use may be prepared according to any method known to the artfor the manufacture of pharmaceutical compositions and such compositionsmay contain one or more agents including sweetening agents, flavoringagents, coloring agents and preserving agents, in order to provide apalatable preparation. Tablets containing the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipient which aresuitable for manufacture of tablets are acceptable. These excipients maybe, for example, inert diluents, such as calcium or sodium carbonate,lactose, lactose monohydrate, croscarmellose sodium, povidone, calciumor sodium phosphate; granulating and disintegrating agents, such asmaize starch, or alginic acid; binding agents, such as cellulose,microcrystalline cellulose, starch, gelatin or acacia; and lubricatingagents, such as magnesium stearate, stearic acid or talc. Tablets may beuncoated or may be coated by known techniques includingmicroencapsulation to delay disintegration and adsorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonostearate or glyceryl distearate alone or with a wax may be employed.

In some embodiments, disclosed herein are oral dosage forms (e.g.,tablets), which may be prepared from hot melt extrusion or spray-dryingdispersion (SDD) technologies.

In some embodiments, disclosed herein are hard capsules filled withpowder, beads, or granules containing the active ingredient in admixturewith non-toxic pharmaceutically acceptable excipient which are suitablefor manufacture of hard or soft capsules. These excipients may be, forexample, inert diluents, such as calcium or sodium carbonate, lactose,lactose monohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc.

In some embodiments, disclosed herein are hard or soft capsules filledwith liquid or semi-solid mixtures containing the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipient which aresuitable for manufacture of hard or soft capsules. These excipients maybe, for example, solubilizing oils such as maize oil, sesame oil, orcorn oil; medium chain triglycerides and related esters, such as,derivitized palm kernel oil or coconut oil; self-emulsifying lipidsystems (SEDDS or SMEDDS), such as caprylic triglyceride or propyleneglycol monocaprylate; viscosity modifiers, such as, cetyl alcohol,steryl alcohol, glycerol stearate; and solubilizing agents andsurfactants, such as polyethylene glycol, propylene glycol, glycerin,ethanol, polyethoxylated castor oil, poloxamers, or polysorbates.

The pharmaceutical compositions of the present disclosure may be in theform of a sterile injectable preparation, such as a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents which have been mentioned herein. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

In some embodiments, the sterile injectable preparation disclosed hereinmay also be a sterile injectable solution or suspension prepared from areconstituted lyophilized powder in a non-toxic parenterally acceptablediluent or solvent, such as a solution in 1,3-butane-diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile fixed oils may conventionally be employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid may likewise be used in the preparation of injectables.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. In certain embodiments the suspension is a microsuspension. Incertain embodiments the suspension is a nanosuspension.

In some embodiments, formulations suitable for parenteral administration(e.g., intramuscular (IM) and subcutaneous (SC) administration) willinclude one or more excipients. Excipients should be compatible with theother ingredients of the formulation and physiologically innocuous tothe recipient thereof. Examples of suitable excipients are well known tothe person skilled in the art of parenteral formulation and may be founde.g., in Handbook of Pharmaceutical Excipients (eds. Rowe, Sheskey &Quinn), 6th edition 2009.

Examples of solubilizing excipients in a parenteral formulation (e.g.,an SC or IM formulation) include, but are not limited to, polysorbates(such as polysorbate 20 or 80) and poloxamers (such as poloxamer 338,188, or 207). In some embodiments, disclosed herein is a parenteraladministration (e.g., an SC or IM formulation) that comprises a cholinesalt of Compound 1, or crystalline form thereof, disclosed herein and apoloxamer, in particular poloxamer 338. In some embodiments, the amountof poloxamer (e.g., poloxamer 388) in a parenteral administrationdisclosed herein is less than about 5%, such as less than about 3%,about 2%, about 1%, or about 0.5%.

In some embodiments, the parenteral formulation (e.g., an SC or IMformulation) disclosed herein is an aqueous suspension. In someembodiments, the parenteral formulation (e.g., an SC or IM formulation)disclosed herein is an aqueous suspension that comprises a choline saltof Compound 1, or crystalline form thereof, disclosed herein and saline.In some embodiments, the parenteral formulation (e.g., an SC or IMformulation) disclosed herein is an aqueous suspension that comprises acholine salt of Compound 1, or crystalline form thereof, disclosedherein saline, and a poloxamer (such as poloxamer 338, 188, or 207).

In certain embodiments, the composition is disclosed as a solid dosageform, including a solid injectable dosage form, such as a solid depotform.

The amount of active ingredient that may be combined with the inactiveingredients to produce a dosage form may vary depending upon theintended treatment subject and the particular mode of administration.For example, in some embodiments, a dosage form for oral administrationto humans may contain approximately 1 to 1000 mg of active materialformulated with an appropriate and convenient amount of carrier material(e.g., inactive ingredient or excipient material). In certainembodiments, the carrier material varies from about 5 to about 95% ofthe total compositions (weight:weight).

It should be understood that in addition to the ingredients particularlymentioned above the compositions of these embodiments may include otheragents conventional in the art having regard to the type of compositionin question, for example those suitable for oral administration mayinclude flavoring agents.

In certain embodiments, a composition comprising an active ingredientdisclosed herein in one variation does not contain an agent that affectsthe rate at which the active ingredient is metabolized. Thus, it isunderstood that compositions comprising a choline salt of Compound 1, orcrystalline form thereof, in certain embodiments do not comprise anagent that would affect (e.g., slow, hinder or retard) the metabolism ofthe choline salt of Compound 1, or crystalline form thereof, or anyother active ingredient administered separately, sequentially orsimultaneously with the salt or crystalline form. It is also understoodthat any of the methods, kits, articles of manufacture and the likedetailed herein in certain embodiments do not comprise an agent thatwould affect (e.g., slow, hinder or retard) the metabolism of a cholinesalt of Compound 1, or crystalline form thereof, or any other activeingredient administered separately, sequentially or simultaneously withthe choline salt of Compound 1, or crystalline form thereof.

Kits and Articles of Manufacture

The present disclosure relates to a kit comprising a choline salt ofCompound 1, or crystalline form thereof, disclosed herein. In oneembodiment, the kit may comprise one or more additional therapeuticagents as described hereinbefore. The kit may further compriseinstructions for use, e.g., for use in inhibiting an HIV reversetranscriptase, such as for use in treating an HIV infection or AIDS oras a research tool. The instructions for use are generally writteninstructions, although electronic storage media (e.g., magnetic disketteor optical disk) containing instructions are also acceptable.

The present disclosure also relates to a pharmaceutical kit comprisingone or more containers comprising a choline salt of Compound 1, orcrystalline form thereof, disclosed herein. Optionally associated withsuch container(s) can be a notice in the form prescribed by agovernmental agency regulating the manufacture, use or sale ofpharmaceuticals, which notice reflects approval by the agency for themanufacture, use or sale for human administration. Each component (ifthere is more than one component) can be packaged in separate containersor some components can be combined in one container wherecross-reactivity and shelf life permit. The kits may be in unit dosageforms, bulk packages (e.g., multi-dose packages) or sub-unit doses. Kitsmay also include multiple unit doses of the compounds and instructionsfor use and be packaged in quantities sufficient for storage and use inpharmacies (e.g., hospital pharmacies and compounding pharmacies).

In some embodiments, the present disclosure also relates to apharmaceutical kit comprising one or more containers comprising acholine salt of Compound 1, or crystalline form thereof, disclosedherein. Optionally associated with such container(s) can be a notice inthe form prescribed by a governmental agency regulating the manufacture,use or sale of pharmaceuticals, which notice reflects approval by theagency for the manufacture, use or sale for human administration. Eachcomponent (if there is more than one component) can be packaged inseparate containers or some components can be combined in one containerwhere cross-reactivity and shelf life permit. The kits may be in unitdosage forms, bulk packages (e.g., multi-dose packages) or sub-unitdoses. Kits may also include multiple unit doses of the compounds andinstructions for use and be packaged in quantities sufficient forstorage and use in pharmacies (e.g., hospital pharmacies and compoundingpharmacies).

Also disclosed are articles of manufacture comprising a unit dosage of acholine salt of Compound 1, or crystalline form thereof, disclosedherein in suitable packaging for use in the methods described herein.Suitable packaging is known in the art and includes, for example, vials,vessels, ampules, bottles, jars, flexible packaging and the like. Anarticle of manufacture may further be sterilized and/or sealed.

EXAMPLES

General Methods

X-Ray Powder Diffraction (XRPD)

XRPD patterns were collected on a PANanalytical XPERT-PRO diffractometerat ambient conditions under the following experimental settings: 45 KV,40 mA, Kα1=1.5406 Å, scan range 2 to 40°, step size 0.0084 or 0.0167°,measurement time: 5 min.

Differential Scanning calorimetry (DSC)

DSC thermograms were collected on a TA Instruments Q2000 system equippedwith a 50 position auto-sampler. The calibration for energy andtemperature was carried out using certified indium. Typically 1-5 mg ofeach sample, in a pin-holed aluminium pan, was heated at 10° C./min from25° C. to 300° C. A purge of dry nitrogen at 50 mL/min was maintainedover the sample throughout the measurement. The onset of the meltingendotherm was reported as the melting point.

Proton Nuclear Magnetic Resonance (¹H NMR)

¹H NMR spectra were collected on a Varian 400-MR 400 MHz instrument with7620AS sample changer. The default proton parameters are as follows:spectral width: 14 to −2 ppm (6397.4 Hz); relaxation delay: 1 sec;pulse: 45 degrees; acquisition time: 2.049 sec; number of scans orrepetitions: 8; temperature: 25° C. Samples were prepared in dimethylsulfoxide-d6, unless otherwise stated. Off-line analysis was carried outusing MestReNova software.

Intermediate 1. tert-Butyl(S)-(1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

Step 1.(S)—N-((3,6-Dibromopyridin-2-yl)methylene)-2-methylpropane-2-sulfinamide

3,6-Dibromopicolinaldehyde (76.0 g, 0.287 mol) and(S)-2-methylpropane-2-sulfinamide (36.51 g, 0.301 mol) were combined inNMP (N-methyl-2-pyrrolidone) (200 mL). To the reaction mixture was addedCs₂CO₃ (41.94 g, 0.316 mol) as a solid in one portion. The reactionmixture was stirred for about 2 h then cooled to about 5° C. Water (1.3L) was added to the reaction mixture. The resulting suspension wasstirred for about 1 h, solids isolated by filtration, washed with water(5×100 mL) and dried to provide the title compound. MS (m/z) 368.9[M+H]⁺.

Step 2.(S)—N—((S)-1-(3,6-Dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-methylpropane-2-sulfinamide

A reaction vessel was charged with(S)—N-((3,6-dibromopyridin-2-yl)methylene)-2-methylpropane-2-sulfinamide(65.5 g, 177.95 mmol) followed by DMF (dimethylformamide) (260 mL). Themixture was stirred for about 5 min until homogeneous and the solutionwas cooled to about 8° C. To the reaction mixture was added(3,5-difluorobenzyl)zinc bromide (0.5 M in tetrahydrofuran (THF), 516.04mL) dropwise over about 90 mins. The mixture was stirred for about anadditional 2.5 h. To the reaction mixture, 5% AcOH (acetic acid) inwater (640 mL) was added over about 10 mins followed by CPME(cyclopentyl methyl ether) (320 mL) in one portion. The mixture wasstirred for about 5 mins, warmed to room temperature, and the layerswere separated. The organic layer was washed with 5% AcOH (320 mL) thentreated with 0.5M NaOH (330 mL) and washed with brine. The organic layerwas collected, dried with Na₂SO₄, and filtered. To the crude mixture wasadded MeOH (methanol) (33 mL). To the stirring mixture was addeddropwise 3M HCl in CPME (128 mL) over about 15 mins. After stirring forabout 1 h, the precipitate was removed by filtration. The filtrate wasdiluted with hexane (300 mL) and the product was extracted with water(450 mL). The aqueous layer was basified with 8M NaOH and extracted withCPME (375 mL). The organic layer was washed with brine, dried overNa₂SO₄ and filtered to provide the title compound in solution which wasused directly in the next reaction. MS (m/z) 497.0 [M+H]⁺.

Step 3.(S)-1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethan-1-amine

The resulting solution of(S)—N—((S)-1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenypethyl)-2-methylpropane-2-sulfinamidewas diluted with CPME to a volume of 700 mL to which acetonitrile (350mL) was added. To the stirring mixture, concentrated HCl (37%, 16.4 mL)was added dropwise over about 10 mins at room temperature. The thickslurry was vigorously stirred for about 4 h. The solids were filteredand washed with 2:1 CPME (cyclopropyl methyl ether):ACN to provide thetitle compound. MS (m/z) 393.3 [M+H]⁺.

Step 4. tert-Butyl(S)-(1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

A reaction vessel was charged with 2-MeTHF (2-methyltetrahydrofuran)(190 mL), water (190 mL) and(S)-1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethan-1-amine(46.9 g, 0.11 mol) followed by portionwise addition of NaHCO₃ (30.34 g,0.36 mol). The reaction mixture was cooled to about 5° C. anddi-tert-butyl dicarbonate (27.47 g, 0.13 mol) was added. The reactionmixture was stirred at about 0° C. for about 2 h and ambient temperaturefor about 2 h. The reaction mixture was diluted with water and extractedwith MTBE (methyl tert-butyl ether). The organic layers were washed withbrine, dried and concentrated. Crude compound was purified by columnchromatography on silica to provide the title compound. MS (m/z) 492.8[M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄) δ 7.85 (d, 1H), 7.42 (d, 1H),6.90-6.72 (m, 3H), 5.33 (dd, 1H), 3.10 (dd, 1H), 2.92 (dd, 1H), 1.36 (s,9H).

Intermediate 2.4-Chloro-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indazol-3-amine

Step 1. 7-Bromo-4-chloro-1H-indazol-3-amine

To 3-bromo-6-chloro-2-fluorobenzonitrile (13.9 g, 59.3 mmol) in EtOH(ethanol) (60 mL) was added hydrazine monohydrate (5.77 mL). Thereaction mixture was heated to about 80° C. for about 3 h. After coolingto ambient temperature, EtOH (20 mL) was added to allow for stirring.The solids were isolated by filtration, washed with cold EtOH, and driedto provide the title compound. MS (m/z) 247.9 [M+H]⁺.

Step 2. 7-Bromo-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-3-amine

A reactor was charged with 7-bromo-4-chloro-1H-indazol-3-amine (397.2 g,1.6 mol) and Cs₂CO₃ (1052 g, 3.2 mol) then diluted with DMF(dimethylformamide) (4000 mL). To this was slowly added2,2,2-trifluoroethyl trifluoromethanesulfonate (463.2 g, 1.9 mol) viaaddition funnel. Upon completion of the addition, the reaction mixturewas allowed to stir for about 1 hour, at which time, H₂O (16 L) wasadded slowly. Upon completion of the addition, the mixture was allowedto stir for about 12 hours at about 15° C. The slurry was filtered andthe collected solids were suspended in DMF (800 mL). To this was addedH₂O (4800 mL) and the resulting solids were collected by filtration anddried to provide the title compound. MS (m/z) 330.1 [M+H]⁺.

Step 3.4-Chloro-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indazol-3-amine

A reaction vessel was charged with7-bromo-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-3-amine (15.00 g,45.66 mmol), bis(pinacolato)diboron (17.39 g, 68.49 mmol), potassiumpropionate (15.36 g, 136.98 mmol), dioxane (90 mL) and DMF(dimethylformamide) (30 mL). Bis(triphenylphosphine)palladium(II)dichloride (0.64 g, 0.91 mmol) was added and the reaction solutiondegassed by bubbling argon for about 2 min. The reaction mixture washeated to about 105° C. for about 4 hrs. After cooling to ambienttemperature, the reaction mixture was filtered through a pad of Celiteand silica gel washing with EtOAc. The filtrate was washed with 5% LiClsolution and brine. The organic layers were separated, dried, andconcentrated under reduced pressure. The residue was treated withIPAc/heptane (1/10) at about 60° C. then cooled to ambient temperatureand stirred for about 15 h. The solids were collected by filtration anddried to afford the title compound. MS (m/z) 376.7 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 7.69 (d, 1H), 7.06 (d, 1H), 5.55 (s, 2H), 5.45 (q, 2H),1.32 (s, 12H).

Intermediate 3.2-((3bS,4aR)-5,5-Difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid

Step 1. Lithium2,2,2-trifluoro-1-(3-oxobicyclo[3.1.0]hexan-2-ylidene)ethan-1-olate

A reactor was charged with bicyclo[3.1.0]hexan-3-one (95.6 g, 0.99 mol)and ethyl 2,2,2-trifluoroacetate (113.2 mL, 0.95 mol) and THF (50 mL).The reaction mixture was cooled to about 0° C. LiHMDS (Lithiumbis(trimethylsilyl)amide) (1 L of 1.0M solution in THF, 1 mol) was addedvia an addition funnel at a rate to maintain internal temperature atbelow about 1° C. After the addition was complete, hexanes (235 mL) wasadded in a steady stream via an addition funnel and stirred for about 15min. The resultant solids were collected by filtration, washed withhexanes (3×400 mL), and dried to provide the title compound.

Step 2. Ethyl2-(3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetate

A reactor was charged with lithium2,2,2-trifluoro-1-(3-oxobicyclo[3.1.0]hexan-2-ylidene)ethan-1-olate(177.2 g, 0.89 mol) and EtOH (ethanol) (779 mL). The temperature wasbrought to and maintained at about 0° C. HCl in dioxane (4.0 N, 443 mL)was added via an addition funnel followed by the addition of solid ethylhydrazinoacetate HCl salt (138.4 g, 0.90 mol). The reaction temperaturewas adjusted to about 35° C. After about 1 h, the reaction volume wasreduced by ˜40% by distillation at reduced pressure. Water (1.3 L) wasadded with vigorous agitation and temperature adjusted to about 15° C.The resultant solids were collected by filtration, washed with water(3×500 mL), hexanes (3×400 mL), and dried to provide the title compound.MS (m/z) 275.1 [M+H]⁺.

Step 3. Ethyl2-(5-oxo-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2]-c]pyrazol-1-yl)acetate

A reactor was charged with ethyl2-(3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetate(291.2 g, 1.06 mol), I (acetonitrile) (1.65 L) and water (825 mL) towhich N-hydroxyphthalimide (17.4 g, 0.103 mol) and NaClO₂ (41.0 g, 0.45mol, ˜20% of total amount to be added) were added. The reaction mixturewas heated to about 50° C. and the remaining NaClO₂ (163.0 g, 1.80 mol)was added in five portions over about 2 h. After consumption of startingmaterial, the temperature was adjusted to about 20° C. and aqueoussodium bisulfite (40% w/w, 350 mL) was added via an addition funnel.Ethyl acetate (1.75 L) was added and the layers were separated. Theaqueous layer was back extracted with EtOAc (ethyl acetate) (500 mL).The organic layers were combined and washed with saturated aqueousNaHCO₃ (500 mL) and 1:1 water/brine (500 mL). The organic layer wasconcentrated under reduced pressure and co-evaporated with IPAc(isopropyl acetate) (300 mL). The crude solid was crystallized from amixture of IPAc/heptane. The resultant solids were collected byfiltration, washed with heptane, and dried to provide the titlecompound. MS (m/z) 289.0 [M+H]⁺.

Step 4.2-(5-Oxo-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid

To a solution of ethyl2-(5-oxo-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetate(80.40 g, 278.95 mmol) in 2-MeTHF (167 mL) was added 2M aqueous sodiumhydroxide (167 mL). After about 25 minutes of stirring at roomtemperature, the reaction mixture was diluted with 2-MeTHF and wasslowly acidified by the dropwise addition of concentrated HCl. Theorganic layer was isolated and the aqueous layer was extracted with anadditional portion of 2-MeTHF. The combined organic layers were washedwith saturated aqueous sodium chloride, then dried over sodium sulfate,filtered, and concentrated. The resulting oil was taken in ethylacetate. Hexanes was added with vigorous stirring until solid formationwas observed. The solid was isolated by filtration and dried to providethe title compound. MS (m/z) 259.00 [M−H]⁻.

Step 5.2-(3-(Trifluoromethyl)-4,4a-dihydrospiro[cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-5,2′[1,3]dithiolane]-1(3bH)-yl)aceticacid

To a solution of2-(5-oxo-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid (3.0 g, 11.5 mmol) in DCM (dichloromethane) (25 mL) was added1,2-ethanedithiol (1.07 mL, 12.68 mmol) followed by borontrifluoride-acetic acid complex (4.0 mL, 28.8 mmol). The reactionmixture was stirred at room temperature overnight. To the reactionmixture was added water (60 mL) and 2-MeTHF (60 mL). The organic layerwas isolated, dried over sodium sulfate, filtered, and concentrated. Thecrude was dissolved in ethyl acetate (2 mL) and the solution dilutedwith hexanes (12 mL) with vigorous stirring to provide a solid. Thesolid was isolated by filtration and dried to provide the titlecompound. MS (m/z) 337.12 [M+H]⁺.

Step 6.2-(5,5-Difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid

To a suspension of 1,3-dibromo-5,5-dimethylhydantoin (12.75 g, 44.6mmol) in DCM (35 mL) was added pyridine hydrofluoride (5.0 mL) at about0° C. The suspension was stirred at about 0° C. for about 10 minutes. Tothe suspension was added a solution of2-(3-(trifluoromethyl)-4,4a-dihydrospiro[cyclopropa[3,4]cyclopenta[1,2-c]pyrazole-5,2′-[1,3]dithiolane]-1(3bH)-yl)aceticacid (5.00 g, 14.9 mmol) dropwise. After addition was complete, thereaction mixture was stirred at about 0° C. for about an additional 15minutes. The reaction mixture was poured into saturated aqueous sodiumbicarbonate solution (300 mL) with vigorous stirring. The organic layerwas removed and the aqueous layer was acidified to pH˜1 withconcentrated HCl. The aqueous phase was extracted with three portions ofMTBE (methyl tert-butyl ether). The combined organic layers were driedover sodium sulfate, filtered, and concentrated. The resulting solid wastaken in MTBE (16 mL) and filtered to remove any resulting solid. Thesolution was then extracted with 2N NaOH (16 mL). The aqueous layer wasdiluted with water (16 mL) with vigorous stirred and stirred at roomtemperature for about 15 minutes. The resulting solid was removed byfiltration. The aqueous layer was acidified by slow, dropwise additionof concentrated HCl to pH˜1 with vigorous stirring to provide a solidprecipitate. The solid was isolated by filtration to provide the titlecompound. MS (m/z) 281.12 [M+H]⁺.

Step 7.2-((3bS,4aR)-5,5-Difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid

2-(5,5-Difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid was separated to its constituent enantiomers(2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid (Intermediate 3) and2-((3bR,4aS)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid), by chiral supercritical fluid chromatography (SFC) under thefollowing conditions: Instrument: Thar 350 preparative SFC; Column:ChiralPak IC-10 u, 300×50 mm I.D; Mobile phase: 35% Isopropanol (0.1%NH₃.H₂O) and CO₂; Flow rate: 200 mL/min; Column temperature: 38° C.; UVdetection: 220 nm; Sample preparation: Compound was dissolved inisopropanol to ˜45 mg/mL; Injection: 6.5 mL per injection. AnalyticalSFC [mobile phase: A for CO₂ and B for Isopropanol (0.05% DEA);Gradient: B 20%; A; Flow rate: 2.35 mL/min; Column: Chiralpak IC-3,150×4.6 mm, 3 μm; Wavelength: 254 nm]. The desired isomer,2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid, eluted at t=3.39 min; ¹H NMR (400 MHz, Chloroform-d) δ 4.93 (s,2H), 2.52-2.43 (m, 2H), 1.44-1.38 (m, 1H), 1.15 (m, 1H).

Intermediate 4: 3-methyl-3-(methylsulfonyl)but-1-yne

To a stirred suspension of sodium methanesulfinate (18.47 g, 175.5 mmol)and copper(I) chloride (1.45 g, 14.6 mmol) in DMF (dimethylformamide)(50 mL) was added 3-chloro-3-methylbut-1-yne (15.00 g, 146.3 mmol, 16.4mL) dropwise. The resulting reaction mixture was heated to about 40° C.and stirred for about 16 h. The reaction mixture was cooled to roomtemperature and diluted with EtOAc. The solution was washed with waterand brine. The organic layer was collected and dried over sodiumsulfate, then filtered. The solution was concentrated under vacuum andpurified by silica gel chromatography to provide the title compound. Mp:114.8-115.5° C. ¹H NMR (400 MHz, Chloroform-d) δ 3.04 (s, 3H), 2.58 (s,1H), 1.67 (s, 6H).

Example 1N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide(Compound 1)

Step 1. tert-Butyl(S)-(1-(3-bromo-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

A reactor was charged with tert-butyl(S)-(1-(3,6-dibromopyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate(Intermediate 1, 50.00 g, 101.8 mmol),3-methyl-3-methylsulfonyl-but-1-yne (17.86 g, 122.2 mmol), DMF(dimethylformamide) (90 mL) and Et₃N (trimethylamine) (42.5 mL, 305.4mmol). The reaction mixture was heated to about 50° C.Bis(triphenylphosphine)palladium(II) dichloride (2.14 g, 3.1 mmol) andcopper(I) iodide (0.58 g, 3.1 mmol) were added. After about 30 min, thereaction mixture was diluted with MeCN (acetonitrile) (200 mL) and then7% aq. NH₄Cl (200 mL) was added dropwise. A slurry was formed andadjusted to ambient temperature. After about 3 h, the solids werecollected by filtration. The cake was washed with MeCN/water (1:1, 75mL) twice and MTBE (methyl tert-butyl ether) (75 mL). The solid wasdried to provide the title compound. MS (m/z) 556 [M+H]⁺. ¹H NMR (400MHz, Chloroform-d) δ 7.84 (d, J=8.2 Hz, 1H), 7.29-7.15 (m, 1H),6.70-6.55 (m, 2H), 5.79 (d, J=9.0 Hz, 1H), 5.57-5.45 (m, 1H), 3.21-3.05(m, 4H), 2.99-2.88 (m, 1H), 1.80 (s, 6H), 1.40* (s, 7H), 1.30* (s, 2H).*denotes presence of atropisomers in 4.6:1 ratio.

Step 2. tert-Butyl(S)-(1-(3-(3-amino-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

tert-Butyl(S)-(1-(3-bromo-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenypethyl)carbamate(1000.0 mg, 1.79 mmol),4-chloro-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1H-indazol-3-amine(808.5 mg, 2.15 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (65.6 mg,0.09 mmol), and cesium carbonate (876.7 mg, 2.69 mmol) were charged in around bottom flask and placed under argon. Dioxane (10 mL) and water (2mL) were added, and the suspension was degassed by bubbling argon forabout one minute. After degassing, the reaction flask was fitted with areflux condenser and heated to about 80° C. for overnight. The reactionmixture was cooled to room temperature, and the aqueous layer wasremoved. The organic layer was concentrated under vacuum, and theresulting residue was purified by silica gel column chromatography toprovide the title compound. MS (m/z) 726.1 [M+H]⁺. ¹H NMR (400 MHz,Chloroform-d) δ 7.69-7.55 (m), 7.55-7.42 (m), 7.16-7.06 (m), 7.07-6.96(m), 6.89 (d), 6.60 (tt), 6.44 (dd), 6.20 (d), 6.16 (d), 6.08 (s),5.69-5.53 (m), 5.29 (s), 5.26 (d), 4.95-4.85 (m), 4.64 (q), 4.59-4.46(m), 4.36-4.19 (m), 3.94-3.76 (m), 3.64-3.54 (m), 3.18 (s), 3.17 (s),3.01-2.84 (m), 2.78-2.68 (m), 1.86-1.82 (m), 1.38 (s), 1.34 (s), 1.26(s), 1.23 (s), 1.15 (s).

Step 3. tert-Butyl(S)-(1-(3-(4-chloro-3-(N-(methylsulfonyl)methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate

tert-Butyl(S)-(1-(3-(3-amino-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate(37.89 g, 52.18 mmol) was dissolved in methylene chloride (380 mL) withstirring at ambient temperature. To it was added triethylamine (21.82mL, 156.54 mmol) followed by slow addition of methanesulfonyl chloride(8.08 mL, 104.36 mmol). When the reaction was complete, water (200 mL)was added and stirred for about 0.5 hours. The organic layer wasseparated and the aqueous layer was extracted with methylene chlorideonce. The combined organic layers were washed with water and brine,dried over MgSO₄, filtered and concentrated to a small volume. Hexaneswas added. The liquid suspension was decanted. The remaining solid wasdried under reduced pressure to afford the title compound. MS (m/z):882.69 [M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄) δ 7.87 (d), 7.83 (d), 7.76(s), 7.74 (s), 7.69 (s), 7.67 (s), 7.65 (s), 7.52-7.47 (m), 7.46 (s),7.37 (d), 7.33 (d), 7.11-7.03 (m), 4.79-4.55 (m), 4.51 (t), 4.36 (dt),4.20-4.05 (m), 3.64 (s), 3.62 (s), 3.60 (s), 3.59 (s), 3.23 (s), 3.04(d), 3.01 (d), 2.95-2.83 (m), 1.81 (s), 1.34 (s), 1.29 (s), 0.98 (s).

Step 4.(S)—N-(7-(2-(1-Amino-2-(3,5-difluorophenyl)ethyl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-3-yl)-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-3-yl)-N-(methylsulfonyl)methanesulfonamide

To tert-Butyl(S)-(1-(3-(4-chloro-3-(N-(methylsulfonyl)methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)carbamate(39 g, 44 mmol) dissolved in methylene chloride (120 mL) was addedtrifluoroacetic acid (80 mL). The reaction mixture was stirred atambient temperature for about 50 minutes. The reaction mixture wasdiluted with methylene chloride and slowly poured into ice coldsaturated aqueous NaHCO₃. The organic layer was separated, washed withwater and brine, dried over MgSO₄, filtered and concentrated to drynessto afford the title compound. MS (m/z): 782.84 [M+H]⁺. ¹H NMR (400 MHz,Chloroform-d) δ 7.61 (d), 7.54-7.44 (m), 7.40 (d), 7.33 (d), 7.20 (d),6.66-6.57 (m), 6.44 (d), 6.33 (d), 6.17 (d), 4.64 (s), 3.68 (s), 3.64(s), 3.61 (s), 3.55 (s), 3.19 (s), 3.05 (dd), 2.85-2.72 (m), 1.86 (s),1.62 (s).

Step 5.N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamide

(S)—N-(7-(2-(1-Amino-2-(3,5-difluorophenyl(ethyl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-3-yl)-4-chloro-1-(2,2,2-trifluoroethyl)-1H-indazol-3-yl)-N-(methylsulfonyl)methanesulfonamide(1757 mg, 2.25 mmol),2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)aceticacid (Intermediate 3, 666 mg, 2.36 mmol), and HATU(1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate) (854 mg, 2.25 mmol) were charged in a roundbottom flask and dissolved in DMF (dimethylformamide) (10.0 mL). To thesolution was added N,N-diisopropylethylamine (0.80 mL, 4.49 mmol) at arapid dropwise rate. After addition was complete, the reaction mixturewas stirred at room temperature for about 15 minutes to provide theintermediateN—((S)-1-(3-(4-chloro-3-(N-(methylsulfonyl)methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenypethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamidewhich was not isolated (MS (m/z) 1046.65 [M+H]⁺). To the solution wasadded 2N aq. sodium hydroxide solution (5.0 mL). The mixture was stirredat room temperature for about 30 minutes. The reaction mixture waspartitioned between water and ethyl acetate. The organic layer wascollected and washed with two portions of 5% lithium chloride solutionfollowed by brine. The organic layer was isolated, dried over sodiumsulfate, filtered, and concentrated under vacuum. The resulting residuewas purified by silica gel column chromatography to yield the titlecompound as an amorphous solid. MS (m/z) 968.24 [M+H]⁺. ¹H NMR (400 MHz,Methanol-d₄) δ 7.87-7.57 (m), 7.33-7.09 (m), 6.80-6.70 (m), 6.54 (d),6.47 (d), 6.37-6.19 (m), 5.02-4.94 (m), 4.90-4.70 (m), 4.70-4.51 (m),3.94 (dq), 3.32-3.28 (m), 3.23 (d), 3.07 (dd, J=13.1, 7.6 Hz), 2.93(dd), 2.68-2.35 (m), 1.81 (s), 1.41 (q), 1.12-1.00 (m). ¹⁹F NMR (377MHz, Methanol-d₄) δ −63.65, −71.78 (t), −72.35 (t), −82.75 (dd), −105.70(ddd), −111.73 to −113.10 (m).

Example 2N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamideN,N,N-trimethylethanolammonium (Choline) Salt

100 mg of Compound 1 and 25 mg of choline hydroxide solution (50% inwater, 1 eq.) were stirred in about 1 mL isopropyl alcohol (IPA). Themixture was dried to afford amorphous solids. About 1 mL IPA was addedto the solids, which remained as amorphous solids after stirring forabout 2 hours. 10 μL of water was added, and the solids were mostlydissolved. The solution was held at about 21° C. and it crystallizedover several days. The crystals, which were obtained after filtrationand drying at about 50° C. under vacuum, were designated as theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm I. ¹H-NMR showed that the N,N,N-trimethylethanolammonium (choline)salt of Compound 1, crystalline Form I was enriched in Isomer B, asshown in FIG. 1 (top trace).

The crystals from the experiment described above were initially bulkycrystals. After two months at ambient conditions in a sealed vialwithout agitation, the crystals became needles. XRPD showed a differentpattern for the needles, when compared to the original bulky crystals,as shown in FIG. 2.

1H-NMR spectrum confirmed the needle crystals were still the cholinesalt, but enriched in Isomer A (FIG. 1, bottom trace). UPLC (UltraPerformance Liquid Chromatography) analysis showed that it containsabout 64.8% Isomer A and about 35.1% Isomer B. After drying, thismaterial lost crystallinity, as shown in FIG. 3 and was designated asthe N,N,N-trimethylethanolammonium (choline) salt of Compound 1,Material A. Additional experiments were conducted by stirring either theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm I or Material A in other solvents at about 22° C. The salts formedfrom MeOH, DCM, or toluene were enriched in Isomer B and the salts fromMeCN, 2-MeTHF, i-PrOAc, EtOH, THF, or MTBE were enriched in Isomer A.

Form I

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form I was enriched in Isomer B and were prepared asdescribed above. The XRPD pattern of the N,N,N-trimethylethanolammonium(choline) salt of Compound 1, crystalline Form I is shown in FIG. 4. Alist of 2-theta peaks is provided in Table 1 below.

TABLE 1 Peak No. 2-theta Rel. Int. (%) 1 5.5 7 2 7.5 41 3 7.9 23 4 9.816 5 13.1 12 6 14.2 11 7 14.9 100 8 15.7 73 9 16.0 45 10 16.8 68 11 17.678 12 19.3 46 13 20.8 35 14 21.6 36 15 22.4 57 16 23.6 25 17 26.8 12

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form I is shown in FIG. 5 and exhibited amelting onset of about 157° C.

Form II

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form II was enriched in Isomer B and was obtained byslurrying the N,N,N-trimethylethanolammonium (choline) salt of Compound1, crystalline Form I in MeOH/IPE (1:1 mixture by volume) for one weekat about 22° C., filtering, and drying the solids at about 22 to about50° C. under vacuum.

The XRPD pattern of the N,N,N-trimethylethanolammonium (choline) salt ofCompound 1, crystalline Form II is shown in FIG. 6. A list of 2-thetapeaks is provided in Table 2 below.

TABLE 2 Peak No. 2-theta Rel. Int. (%) 1 7.5 16 2 9.6 15 3 14.0 11 414.7 36 5 14.9 43 6 16.1 43 7 16.9 100 8 18.5 17 9 19.1 21 10 19.6 19 1120.2 22 12 20.8 56 13 21.0 36 14 22.5 18 15 23.3 16 16 24.5 20 17 25.411 18 26.5 28 19 27.8 18 20 30.7 13 21 33.0 6 22 34.1 7 23 35.2 11

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form II is shown in FIG. 7 and exhibited amelting onset of about 147° C.

Form III

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form III was enriched in Isomer B and obtained by slurryingthe N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form I or Material A in DCM/heptane mixture (1:1 mixture byvolume) at about 22° C. for about 24 h, filtering, and drying the solidsat about 50° C. under vacuum.

The XRPD pattern of the N,N,N-trimethylethanolammonium (choline) salt ofCompound 1, crystalline Form III is shown in FIG. 8. A list of 2-thetapeaks is provided Table 3 below.

TABLE 3 Peak No. 2-theta Rel. Int. (%) 1 7.8 30 2 8.1 29 3 8.3 23 4 9.816 5 11.4 8 6 12.3 7 7 13.6 4 8 14.1 15 9 15.0 36 10 15.7 100 11 16.2 3112 16.7 46 13 17.6 35 14 18.1 31 15 19.2 28 16 19.4 30 17 20.0 53 1821.1 37 19 21.7 53 20 22.3 17 21 23.0 26 22 23.7 20 23 24.8 8 24 26.9 10

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form III is shown in FIG. 9 and exhibited amelting onset of about 144° C.

Form IV

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form IV is enriched in Isomer B and was obtained byslurrying the N,N,N-trimethylethanolammonium (choline) salt of Compound1, crystalline Form I or Material A in toluene for about 2 weeks atabout 22° C., filtering, and drying the solids at about 50° C. undervacuum. The XRPD pattern of the N,N,N-trimethylethanolammonium (choline)salt of Compound 1, crystalline Form IV is shown in FIG. 10. A list of2-theta peaks is provided in Table 4 below.

TABLE 4 Peak No. 2-theta Rel. Int. (%) 1 7.5 20 2 8.0 24 3 9.5 12 4 9.910 5 11.9 7 6 14.0 11 7 14.8 57 8 16.1 19 9 17.0 100 10 18.6 11 11 19.721 12 20.3 72 13 21.1 44 14 21.7 22 15 22.3 25 16 23.7 23 17 24.6 21 1825.6 9 19 26.7 29 20 27.7 12 21 29.6 7 22 30.9 12

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form IV is shown in FIG. 11 and exhibited amelting onset of about 136° C.

Form V

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1, Form Vwas enriched in Isomer A and was prepared by slurrying theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, Material Ain MeCN/IPE (1:1 mixture by volume) at about 22° C., filtering, anddrying the solids at about 50° C. under vacuum.

The XRPD pattern of the N,N,N-trimethylethanolammonium (choline) salt ofCompound 1, crystalline Form V is shown in FIG. 12. A list of 2-thetapeaks is provided in Table 5 below.

TABLE 5 Peak No. 2-theta Rel. Int. (%) 1 6.9 28 2 7.9 54 3 10.7 52 411.9 15 5 12.5 31 6 12.9 19 7 14.0 20 8 14.5 49 9 14.8 34 10 15.6 43 1115.9 61 12 16.7 53 13 17.0 45 14 17.3 53 15 17.6 100 16 17.8 62 17 18.728 18 19.1 22 19 19.6 18 20 21.1 65 21 21.8 59 22 22.8 62 23 23.2 45 2423.9 33 25 24.4 6 26 25.1 10 27 25.7 24 28 26.0 29 29 26.9 48 30 27.3 2931 28.1 9 32 29.1 26 33 31.4 11 34 31.7 22 35 35.0 12

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form V is shown in FIG. 13 and exhibited amelting onset of about 159° C.

Form VI

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form VI was enriched in Isomer A and was prepared byslurrying the N,N,N-trimethylethanolammonium (choline) salt of Compound1, Material A in 2-MeTHF at about 22° C. for about 24 h, filtering, anddrying the solids at about 50° C. under vacuum.

The XRPD pattern of the N,N,N-trimethylethanolammonium (choline) salt ofCompound 1, crystalline Form VI is shown in FIG. 14. A list of 2-thetapeaks is provided in Table 6 below.

TABLE 6 Peak No. 2-theta Rel. Int. (%) 1 4.2 5 2 6.1 21 3 8.6 63 4 9.526 5 11.7 9 6 13.8 14 7 15.4 20 8 16.6 18 9 18.3 25 10 19.1 28 11 20.4100 12 21.9 53 13 22.5 55 14 24.2 37 15 25.2 34 16 28.3 9

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, Form VI is shown in FIG. 15 and exhibited a melting onsetof about 121° C.

Form VII

The N,N,N-trimethylethanolammonium (choline) salt of Compound 1,crystalline Form VII was enriched in Isomer A and was prepared byslurrying the N,N,N-trimethylethanolammonium (choline) salt of Compound1, Material A in i-PrOAc at about 22° C. for about 24 h, filtering, anddrying the solids at about 50° C. under vacuum.

The XRPD pattern The N,N,N-trimethylethanolammonium (choline) salt ofCompound 1, crystalline Form VII is shown in FIG. 16. A list of 2-thetapeaks is provided in Table 7 below.

TABLE 7 Peak No. 2-theta Rel. Int. (%) 1 4.7 13 2 5.5 3 3 7.3 100 4 8.988 5 9.5 76 6 13.3 13 7 14.0 15 8 15.2 18 9 16.4 15 10 17.6 9 11 18.3 2912 19.6 24 13 20.5 32 14 21.5 27 15 22.3 55 16 23.9 9 17 24.9 36 18 26.58 19 28.4 18 20 30.7 3 21 35.8 3

The DSC thermogram of the N,N,N-trimethylethanolammonium (choline) saltof Compound 1, crystalline Form VII is shown in FIG. 17 and exhibited amelting onset of about 144° C.

Example 3N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamideN,N,N-trimethylethanolammonium (choline) salt, Ethanol Solvate

The ethanol solvate form of the N,N,N-trimethylethanolammonium (choline)salt of Compound 1 was enriched in Isomer A and was prepared byslurrying the N,N,N-trimethylethanolammonium (choline) salt of Compound1, crystalline Form I or Material A in EtOH/heptane mixture (1:1 mixtureby volume) at about 22° C. for about 24 h. The XRPD pattern of theethanol solvate form is shown in FIG. 18. After drying, this materiallost crystallinity.

Example 4N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamideN,N,N-trimethylethanolammonium (choline) salt, Tetrahydrofuran (THF)Solvate

The tetrahydrofuran (THF) solvate form of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1 was enrichedin Isomer A and was prepared by slurrying theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, crystallineForm I or Material A in THF/heptane mixture (1:1 mixture by volume) atabout 22° C. for about 24 h. The XRPD pattern of the tetrahydrofuransolvate form is shown in FIG. 19. After drying, this material lostcrystallinity.

Example 5N—((S)-1-(3-(4-Chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamideN,N,N-trimethylethanolammonium (choline) salt, Methyl tert-Butyl Ether(MTBE) Solvate

The methyl tert-butyl ether (MTBE) solvate form of theN,N,N-trimethylethanolammonium (choline) salt of Compound 1 was enrichedin Isomer A and was prepared by slurrying theN,N,N-trimethylethanolammonium (choline) salt of Compound 1, Material Ain MTBE at about 22° C. for about two weeks. The XRPD pattern of themethyl tert-butyl ether (MTBE) solvate is shown in FIG. 20. Afterdrying, this material lost crystallinity.

Representative synthesis of the compounds described herein can also befound in U.S. patent application Ser. No. 15/680,041, filed Aug. 17,2017, which published as U.S. Patent Application No. 2018-0051005 A1 onFeb. 22, 2018, the entire contents of which are incorporated herein byreference in their entireties.

BIOLOGICAL EXAMPLES Example A

Test A: Antiviral Assay in MT4 Cells

For the antiviral assay, 0.4 μL of 189X test concentration of 3-foldserially diluted compound in DMSO was added to 40 μL of cell growthmedium (RPMI 1640, 10% FBS, 1% Penicillin-Streptomycin, 1% L-Glutamine,1% HEPES) in each well of 384-well plate (10 concentrations) inquadruplicate.

1 mL Aliquots of MT4 cells were pre-infected for 3 hours at 37° C. with25 μL of cell growth medium (mock-infected) or a fresh 1:250 dilution ofan HIV-IIIb concentrated ABI stock (0.004 m.o.i.). Infected anduninfected cells were diluted in cell growth media and 35 μL (2000cells) was added to each well of the assay plates.

Assay plates were then maintained in a humidified, 5% CO₂ incubator at37° C. After 5 days of incubation, 25 μL of 2× concentratedCellTiter-Glo™ Reagent (catalog # G7573, Promega Biosciences, Inc.,Madison, Wis.) was added to each well of the assay plate. Cell lysis wascarried out by incubating at room temperature for 10 minutes and thenchemiluminescence was read using an Envision plate reader (PerkinElmer).EC₅₀ values were calculated as the compound concentration that caused a50% decrease in luminescence signal, a measure of HIV-1 replication.

As described in Examples B-D, Compound 1 provides advantages compared tostructurally close compounds (herein designated as Compounds A and B)disclosed in U.S. Patent Publication Nos. 2014/0296266A1 and2014/0303164A1:

Example B

Test B: Cytotoxicity Assay

Compound cytotoxicity and the corresponding CC₅₀ values was determinedusing the same protocol as described in the antiviral assay (Test A)except that uninfected cells were used.

The compound of the present disclosure demonstrates antiviral activity(Test A) as depicted in Table A in comparison to Compound A and CompoundB.

TABLE A Compound EC₅₀ (nM) CC₅₀ (nM) Compound 1 0.185 30068 Compound A1.715 21839 Compound B 2.991 14491

Example C

Test C. Pharmacokinetic Analysis Following Intravenous Administration toSprague-Dawley Rats and Beagle Dogs and Cynomologous Monkeys

Test Article and Formulation

Compound 1 IV administration was formulated in 5% ethanol, 20% PG, 45%PEG 300, 30% pH 2 (0.01N HCl) water at 0.5 mg/mL. Compound A andCompound B intravenous infusion doses were formulated in a sterilesolution of 5% ethanol, 45% PEG 400 and 50% water (pH 2.0) at 0.5 mg/mL.All IV formulations were in solution.

Animals Used

Each rat IV dosing group consisted of 3 male SD rats. At dosing, theanimals generally weighed between 0.317 and 0.355 kg. The animals werefasted overnight prior to dose administration and up to 4 hr afterdosing. Each dog IV dosing group consisted of 3 male, naïve beagle dogs.At dosing, the animals weighed ˜10-12 kg. The animals were fastedovernight prior to dose administration and up to 2 hr after dosing.

Each cynomolgus (cyno) monkey IV dosing group consisted of 3 male, naïvecyno monkeys At dosing, the animals weighed ˜3.2-4 kg. The animals werefasted overnight prior to dose administration and up to 2 hr afterdosing.

Dosing

For the IV infusion group, the test compound was administered byintravenous infusion over 30 minutes. The rate of infusion was adjustedaccording to the body weight of each animal to deliver a dose of 1 mg/kgat 2 mL/kg.

Sample Collection

Serial venous blood samples (approximately 0.4 mL each for rat and 1.0mL for dog) were taken at specified time points after dosing from eachanimal. The blood samples were collected into Vacutainer™ tubes(Becton-Disckinson Corp, New Jersey, USA) containing EDTA as theanti-coagulant and were immediately placed on wet ice pendingcentrifugation for plasma. Centrifugation began within 1 hour ofcollection. All samples were placed into 96-well tubes and maintained ondry ice prior to storage at approximately −70° C.

Determination of the Concentrations of Compound 1 in Plasma

An LC/MS/MS method was used to measure the concentration of testcompounds in plasma.

Calculations

Non-compartmental pharmacokinetic analysis was performed on the plasmaconcentration-time data. A summary of pharmacokinetic parameters areshown in Tables B and C below.

TABLE B Rat Rat Rat Dog Dog Dog Cyno Cyno Cyno CL V_(ss) t_(1/2) CLV_(ss) t_(1/2) CL Vss t_(1/2) Compound (L/h/kg) (L/kg) (h) (L/h/kg)(L/kg) (h) (L/h/kg) (L/kg) (h) Compound 0.05 1.8 28 0.07 1.6 22 0.24 2.712 1 Compound 0.50 1.0 2 0.25 0.8 4 0.45 1.18 2.3 A Compound 0.43 1.4 30.28 1.3 6 0.42 1.59 3.4 B CL: observed clearance; Vss: volume ofdistribution at steady state; t_(1/2): terminal half-life

TABLE C Rat Dog Cyno Rat AUC_(inf) Dog AUC_(inf) Cyno AUC_(inf) CompoundC_(max) (μM · h) C_(max) (μM · h) C_(max) (μM · h) Compound 1.8 19 2.214.8 1.3 4.5 1 Compound 1.4 2.7 2.1 5 1.8 2.6 A Compound 1.1 2.7 1.4 4.31.4 2.9 B AUC_(inf): Area Under the Curve from t = 0 to infinity;C_(max): Maximum plasma concentration

Example D

Test D. Metabolic Stability in Cultured Human Liver Hepatocytes

Radiolabelled test compounds, wherein tritium was introduced into thestructure in place of one or more hydrogens, were prepared according toknown methods in the art.

The radiolabelled compounds were incubated in pooled cryopreservedhepatocytes at a substrate concentration of 0.25 μM and radioactivityconcentration of 10 μCi/mL. The final hepatocyte concentration was 1million cells/mL. The hepatocyte/compound reaction mixture was dissolvedin InVitroGRO™ KHB buffer (catalog # Z99074, BioreclamationlVT, Inc.,Baltimore, Md.) at pH 7.4. The incubations were performed in duplicate.A cell free control and a positive control were included in theincubations. The incubations were carried out with gentle shaking in a37° C. incubator under a humid atmosphere of 95% air/5% CO₂ (v/v).Aliquots (100 mL) were removed after 0, 1, 3, and 6 hours and added to200 mL quenching solution that comprised 0.1% (v/v) TFA in 5% water/95%acetonitrile (v/v). The samples were placed on a shaker for 10 min,followed by centrifugation at 3000 g for 30 min. The samples of thesupernatant were analyzed on a Dionex HPLC/PerkinElmer FlowScintillation Analyzer as described below.

Liquid Chromatography-Radiochromatography

Quantification was done by comparison of radiolabeled metabolites andparent peaks measured on a Radiomatic 625TR Flow Scintillation Analyzercoupled to a Dionex/Chromeleon chromatography system. The column was aPhenomenex Synergi fusion RP (150×4.6 mm, 4 mm) maintained at 32 degreesCelsius. Mobile Phase A consisted of 0.1% (v/v) TFA in 99% water/1%acetonitrile (v/v). Mobile Phase B consisted of 0.1% (v/v) TFA in 5%water/95% acetonitrile (v/v). The flow rate was 1 mL/min using a sampleinjection volume of 100 mL. Gradient was as following: Mobile phase Bwas linearly increased from 2% to 75% over 47 min, maintained at 75% for3 min, changed back to 2%, maintained at 2% for 10 min.

Metabolic stability was determined by measuring the change in relativeabundance of metabolites and parent over time and calculating from itthe rate of disappearance of the parent compound. The stability data wasutilized to calculate predicted human hepatic clearance values accordingto methods known in the art. The predicted human hepatic clearancevalues are shown in Table D below.

TABLE D Predicted Human Hepatic Clearance (L/hr/kg) Compound 1 0.01Compound A 0.09 Compound B 0.04The following can be deduced from the above comparative data:

Compound 1 is more potent in an HIV antiviral assay relative tocompounds A and B (about 9 and about 16 times more potent,respectively). Compound 1 has a longer in vivo terminal half-life in ratrelative to compounds A and B (about 14 and about 9 times longer,respectively). Compound 1 has a lower in vivo clearance in rat relativeto compounds A and B (about 10 and about 8.6 times lower, respectively).Compound 1 has a longer in vivo terminal half-life in dog relative tocompounds A and B (about 5 and about 4 times longer, respectively).Compound 1 has a lower in vivo clearance in dog relative to compounds Aand B (about 3 and about 4 times lower, respectively). Compound 1 ismore stable in human hepatocytes with a lower predicted hepaticclearance relative to compounds A and B (about 9 and about 4 times morestable, respectively).

The above data demonstrate that Compound 1, has improved antiviralpotency and an improved pharmacokinetic profile (which is demonstratedby longer half-life in rat and dog and lower predicted human clearance)when compared to compounds A and B.

The specific pharmacological responses observed may vary according toand depending on the particular active compound selected or whetherthere are present pharmaceutical carriers, as well as the type offormulation and mode of administration employed, and such expectedvariations or differences in the results are contemplated in accordancewith practice of the present disclosure.

The Examples disclosed herein describe the synthesis of compounds,salts, and crystalline forms disclosed herein as well as intermediatesused to prepare the compounds. It is to be understood that individualsteps described herein may be combined. It is also to be understood thatseparate batches of a compound may be combined and then carried forth inthe next synthetic step.

All references, including publications, patents, and patent documentsare incorporated by reference herein, as though individuallyincorporated by reference. The present disclosure provides reference tovarious embodiments and techniques. However, it should be understoodthat many variations and modifications may be made while remainingwithin the spirit and scope of the present disclosure.

What is claimed is:
 1. A pharmaceutically acceptable salt, which isN—((S)-1-(3-(4-chloro-3-(methylsulfonamido)-1-(2,2,2-trifluoroethyl)-1H-indazol-7-yl)-6-(3-methyl-3-(methylsulfonyl)but-1-yn-1-yl)pyridin-2-yl)-2-(3,5-difluorophenyl)ethyl)-2-((3bS,4aR)-5,5-difluoro-3-(trifluoromethyl)-3b,4,4a,5-tetrahydro-1H-cyclopropa[3,4]cyclopenta[1,2-c]pyrazol-1-yl)acetamideN,N,N-trimethylethanolammonium salt.
 2. A crystalline form of the saltof claim
 1. 3. The crystalline form of claim 2, which is crystallineForm I.
 4. The crystalline form of claim 3, wherein the crystalline FormI has at least three XRPD peaks, in terms of 2-theta±0.2°, selected from5.5°, 7.5°, 7.9°, 14.9°, 15.7°, 16.8°, 17.6°, 19.3°, and 22.4°.
 5. Thecrystalline form of claim 3, wherein the crystalline Form I ischaracterized by an XRPD pattern substantially as shown in FIG.
 4. 6.The crystalline form of claim 3, wherein the crystalline Form I ischaracterized by a DSC thermogram having a melting onset of about 157°C.
 7. The crystalline form of claim 3, wherein the crystalline Form I ischaracterized by a DSC thermogram substantially as shown in FIG.
 5. 8.The crystalline form of claim 2, which is crystalline Form II.
 9. Thecrystalline form of claim 8, wherein the crystalline Form II has atleast three XRPD peaks, in terms of 2-theta±0.2°, selected from 7.5°,9.6°, 14.0°, 14.9°, 16.1°, 16.9°, 20.8°, 21.0°, and 26.5°.
 10. Thecrystalline form of claim 8, wherein the crystalline Form II ischaracterized by an XRPD pattern substantially as shown in FIG.
 6. 11.The crystalline form of claim 8, wherein the crystalline Form II ischaracterized by a DSC thermogram having a melting onset of about 147°C.
 12. The crystalline form of claim 8, wherein the crystalline Form IIis characterized by a DSC thermogram substantially as shown in FIG. 7.13. The crystalline form of claim 2, which is crystalline Form III. 14.The crystalline form of claim 13 wherein the crystalline Form III has atleast three XRPD peaks, in terms of 2-theta±0.2°, selected from 7.8°,8.1°, 8.3°, 15.0°, 15.7°, 16.7°, 20.0°, 21.1°, and 21.7°.
 15. Thecrystalline form of claim 13, wherein the crystalline Form III ischaracterized by an XRPD pattern substantially as shown in FIG.
 8. 16.The crystalline form of claim 13, wherein the crystalline Form III ischaracterized by a DSC thermogram having a melting onset of about 144°C.
 17. The crystalline form of claim 13, wherein the crystalline FormIII is characterized by a DSC thermogram substantially as shown in FIG.9.
 18. A pharmaceutical composition comprising the pharmaceuticallyacceptable salt of claim 1, and at least one pharmaceutically acceptableexcipient.
 19. A pharmaceutical composition comprising the crystallineform of claim 2, and at least one pharmaceutically acceptable excipient.20. A pharmaceutical composition comprising the crystalline form ofclaim 3, and at least one pharmaceutically acceptable excipient.
 21. Apharmaceutical composition comprising the crystalline form of claim 8,and at least one pharmaceutically acceptable excipient.
 22. Apharmaceutical composition comprising the crystalline form of claim 13,and at least one pharmaceutically acceptable excipient.